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WO2010126600A1 - Optimal fret pairs and related methods - Google Patents

Optimal fret pairs and related methods Download PDF

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Publication number
WO2010126600A1
WO2010126600A1 PCT/US2010/001268 US2010001268W WO2010126600A1 WO 2010126600 A1 WO2010126600 A1 WO 2010126600A1 US 2010001268 W US2010001268 W US 2010001268W WO 2010126600 A1 WO2010126600 A1 WO 2010126600A1
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WO
WIPO (PCT)
Prior art keywords
tamra
gln
abu
asn
pro
Prior art date
Application number
PCT/US2010/001268
Other languages
French (fr)
Inventor
Anita Hong
Vera Rakhmanova
Original Assignee
Anaspec, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication of WO2010126600A1 publication Critical patent/WO2010126600A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
    • C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses

Definitions

  • the present invention is generally directed to biological assays. More specifically it is directed to optimal FRET pairs and methods related to their use.
  • Proteases are enzymes that cleave protein substrates to form smaller proteins.
  • the smaller proteins exhibit specific beneficial or detrimental functions in the host organism. Where detrimental functions are exhibited, the proteins oftentimes are the root cause of various diseases. Examples of such diseases include cancer, arthritis, Alzheimer's, hypertension, hepatitis and AIDS.
  • protease-based pharmaceuticals required the emergence of new assays that one could use to monitor the effect of small or large molecules on protease activity.
  • Preferred assays were homogeneous, meaning one could obtain a continuous reading of activity rather than stopping enzymatic activity, separating reaction components and then obtaining an activity reading.
  • Useful assays further had to be very sensitive so one could measure activity even when only a small amount of protease was present in the reaction medium.
  • spectrophotometric- based assays appeared to be the likely answer for probing protease activity; they are homogeneous, fast, accurate and easy to use.
  • Spectrophotometric assays are either fluorogenic or chromogenic, with fluorogenic assays being superior in many cases. This is because the fluorogenic assays have a wider linear dynamic range and offer better reproduceability.
  • Forster resonance energy transfer (“FRET”) assays are a type of fluorogenic spectrophotometric assay pharmaceutical companies have oftentimes used for rapid activity screening.
  • FRET involves the transfer of excited-state energy from one type of compound ("donor") to another type (“acceptor”).
  • the donor typically transfers (i.e., emits) short wavelengths of light that are within the wavelength range the acceptor can absorb.
  • Energy transfer occurs within a specified distance between the donor and acceptor, usually 10 to 100 A. The distance at which energy transfer equals 50 percent is called the Forster radius. Within this distance, a decrease in donor fluorescence can be detected upon donor to acceptor energy transfer.
  • FRET fluorescence resonance energy transfer spectrometry
  • the use of FRET in a protease assay involves the construction of a protease substrate that includes both a donor and an acceptor positioned on opposite sides of the cleavage site at an appropriate distance from one another.
  • the substrate is typically constructed from a peptide sequence derived from a known protein cleaved by the protease.
  • Spectrophotometric methods allow one to observe whether the protease substrate has been cleaved, since cleavage provides that the donor-acceptance distance exceeds the Forster radius; this results in recovery of the donor's fluorescence. Reports that discuss the use of FRET-based assays for examination of protease activity include, for example: U.S. Pat. No.
  • the present invention is generally directed to biological assays. More specifically it is directed to optimal FRET pairs and methods related to their use.
  • the present invention is directed to FRET-based protease substrates selected from the list of substrates shown in the Detailed Description above.
  • the present invention is directed to a method of performing a FRET-based assay, where the assay includes the following steps: adding a solution or suspension containing one or more different proteins to a reaction vessel; adding a liquid comprising a FRET-substrate to the reaction vessel, wherein the FRET substrate is selected from the list of substrates shown in the Detailed Description section below.
  • the present invention is directed to a kit for performing a FRET- based assay, wherein the kit comprises a protease substrate.
  • the protease substrate is selected from the list of substrates shown in the Detailed Description section below.
  • FIGS 1-6 show the structures of various members of optimal FRET pairs according to the present invention. Detailed Description of the Invention
  • a donor/acceptor pair for inclusion within the substrate should be chosen such that the absorption spectrum of the acceptor overlaps with the emission spectrum of the donor.
  • a fluorescent donor and a non-fluorescent acceptor are used to make protease peptide substrates.
  • the donor/acceptor pair choice may be limited by the fiuorometer filter at hand.
  • Use of non-native sequences may be used to increase cleavage efficiency, to protect the peptide from degradation or to increase peptide solubility.
  • an ester linkage can replace an amide linkage in the peptide to increase cleavage efficiency.
  • fluorophores react with amino groups, meaning they can be conjugated to the ⁇ -amino group of Lysine.
  • Thiol reactive dyes can be used to conjugate donors and/or acceptors to Cys-containing peptides. This is an economical way to utilize the donors and/or acceptors since the peptides can be HPLC purified first before reacting with the dyes.
  • Lysines or Arginines may be added to increase solubility. These amino acids must be added at the appropriate positions without adversely affecting the protease recognition site.
  • X 2 E, S, G, V, H, L 5 K, P, Q, Y, Cha, A;
  • X 3 D, Q, Cha, N, P, A, G, S 5 L 5 V, K;
  • X 4 E, N 5 Abu, L 5 F 5 Q 5 1 5 R, G 5 A, K, T 5 P, Nva, C(Me) 5 Y;
  • X 5 Abu-y-[COO] 5 Y 5 Smc, D 5 H, A 5 L, V, P 5 G 5 S, E 5 R 5 Dap, W 5 K;
  • X 6 A 5 P 5 HAr 5 L 5 V 5 F, Q, R 5 S 5 G, E, M 5 H 5 Ar 5 Dab, or no amino acid;
  • X 7 S 5 1 5 K, E, V, R, F, A, P, G, H, Nva, Dap, or no amino acid;
  • X 8 C, V, A, F 5 1 5 S, R 5 E, K, P, S, W, Dab, Q 5 or no amino acid;
  • X 9 Q 5 K 5 H 5 S 5 L 5 A 5 1, K(Ac), D, R, or no amino acid;
  • X 10 K, S, A, I, Y, P, L, G, or no amino acid;
  • X 1 I R, K, A, NIe, L, or no amino acid
  • Xi 2 K, F, or no amino acid
  • Xi 4 T, or no amino acid
  • X 15 E, or no amino acid
  • Xi 6 G, or no amino acid
  • Xi 7 K, or no amino acid.
  • Nonlimiting examples of peptide substrates to which the donor and acceptor are conjugated include ("*" designating point of FRET moiety attachment):
  • HCV NS3/4A protease substrate *-Asp-Glu-Asp-Glu-Glu-Abu-y-[COO]Ala- Ser-Cys(*)-NH 2 (SEQ ID NO. 1).
  • HIV-I protease substrate *- ⁇ -Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(*)-OH (SEQ ID NO. 2).
  • MMP generic substrate *-KGP-Cha-Abu-Smc-HAr-K(*)-AK-NH 2 (SEQ ID NO. 3).
  • Renin substrate ⁇ -KHPFHLVIHK-* (SEQ ID NO. 5).
  • TACE B-secretase substrate: *-PLAQ AVRSSSRK-* (SEQ ID NO. 6).
  • Cathepsin D substrate ⁇ -GKPILFFRLK-* (SEQ ID NO. 7).
  • Cathepsin S substrate * -EKARVL AE AAK-* (SEQ ID NO. 8).
  • Cathepsin K substrate *-HPGGPQK-* (SEQ ID NO. 9).
  • Aggrecanase-1 substrate *-AELQGRPISIAK-* (SEQ ID NO. 10).
  • Aggrecanase-2 substrate * -TESESRGAI YK-* (SEQ ID NO. 11).
  • Calpain substrate A ⁇ -PLLAERPK-* (SEQ ID NO. 12).
  • Calpain substrate B ⁇ -TPLKSPPPSPRK-* (SEQ ID NO. 13).
  • Sirtuin substrate *-GQSTSSHS-K(Ac)-L-Nle-FSTEG-K-* (SEQ ID NO. 14).
  • MMP-I substrate *-PLA-Nva-Dap-* (SEQ ID NO. 16).
  • MMP-2 substrate * -PLGC(Me)HARK-* (SEQ ID NO. 17).
  • MMP-3 substrate * -RPKP VE-Nva- WRK-* (SEQ ID NO. 18).
  • MMP-7 substrate *-P YAYWMRK-* (SEQ ID NO. 19).
  • MMP-8 substrate *-P-Cha-G-Nva-HA-Dap-* (SEQ ID NO. 20).
  • MMP-9 substrate *- ⁇ -Abu-P-Cha-Abu-Smc-HA-Dab-* (SEQ ID NO. 21).
  • MMP-IO substrate * -RPKPL A-Nva- WK-* (SEQ ID NO. 22).
  • MMP- 12 substrate *-PLGLEEAK-* (SEQ ID NO. 23).
  • MMP-13 substrate *-PLGLWArK-* (SEQ ID NO. 24).
  • MMP- 14 substrate *- ⁇ -Abu-PQGL-Dab-* (SEQ ID NO. 25).
  • FRET substrates - i.e., peptide/donor/acceptor combinations - according to the present invention are shown below.
  • “Al” through “A6”, “Bl” through “B6”, “Cl” through “C4" and 5-TAMRA designate moieties of a FRET pair. (See FIGS 1-6.) The moiety is attached through a reaction with an activated carboxylic acid group of an ⁇ , ⁇ - unsaturated sulfone group, with the point of attachment shown as “*”.
  • “5-TAMRA” is a rhodamine dye (i.e., 5-carboxytetramethylrhodamine) and is attached to the substrate using chemistry known to those of skill in the art.
  • Nonlimiting examples of optimal FRET pairs/substrate combinations include:
  • Nonlimiting examples of substrates including Al through A6 and 5-TAMRA FRET pairs include:
  • the same substrates may include optimal FRET pairs selected from Bl through B6 combined with Cl through C4 in a manner similar to the nonlimiting examples of substrates including optimal Al through A6 and 5 -TAMRA FRET pairs shown above.
  • Nonlimiting examples of generic substrates including Al through A6 and 5- TAMRA FRET pairs include (where moieties are defined as above):
  • AI-X I -X 2 -N-G-X 5 -X 6 -X 7 -(S-TAMRA)J AI-X I -X 2 -N-G-X 5 -X 6 -X 7 -(S-TAMRA)J
  • A5-X 1 - X 2 -N-G-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X 1 -X 2 -N-G-X 5 -X 6 -X 7 -(S-TAMRA)J
  • AI-XI-X 2 -G-L-X 5 -X 6 -X 7 -(S-TAMRA)J A ⁇ -X I -X 2 -G-L-X 5 -X 6 -X 7 -(S-TAMRA); A3-X I -X 2 -G-L-X 5 -X 6 -X 7 -(5-TAMRA); A4-X I -X 2 -G-L-X 5 -X 6 -X 7 -(5-TAMRA)J A5-X,- X 2 -G-L-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X 1 -X 2 -G-L-X 5 -X 6 -X 7 -(S-TAMRA); Al-X 1 -X 2 -G-F-X 5 -X 6 -X 7 -(S-TAMRA); A2-X r X 2 -G-F-X 5
  • AI-X 1 -X 2 -G-A-X 5 -X 6 -X 7 -(S-TAMRA)J AZ-X I -X 2 -G-A-X 5 -X 6 -X 7 -(S-TAMRA);
  • A3-X 1 -X 2 -G-A-X 5 -X 6 -X 7 -(5-TAMRA)j A4-X,-X 2 -G-A-X 5 -X 6 -X 7 -(5-TAMRA);
  • A5-X 1 - X 2 -G-A-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X 1 -X 2 -G-A-X 5 -X 6 -X 7 -(S-TAMRA)J
  • AI-X 1 -X 2 -G-Y-X 5 -X 6 -X 7 -(S-TAMRA)J AZ-X 1 -X 2 -G-Y-X 5 -X 6 -X 7 -(S-TAMRA); AS-X 1 -X 2 -G-Y-X 5 -X 6 -X 7 -(S-TAMRA); A4-X I -X 2 -G-Y-X 5 -X 6 -X 7 -(5-TAMRA); A5-X,- X 2 -G-Y-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X I -X 2 -G-Y-X 5 -X 6 -X 7 -(S-TAMRA)J
  • a 1 -X 1 -X 2 -L-K-X 5 -X 6 -X 7 -(S-TAMRA)J A2-X,-X 2 -L-K-X 5 -X 6 -X 7 -(5-TAMRA); A3-Xi-X 2 -L-K-X 5 -X 6 -X 7 -(5-TAMRA)j A4-X,-X 2 -L-K-X 5 -X 6 -X 7 -(5-TAMRA); A5-Xi- X 2 -L-K-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X 1 -X 2 -L-K-X 5 -X 6 -X 7 -(S-TAMRA); Al-Xi-X 2 -L-T-X 5 -X6-X 7 -(5-TAMRA); A2-X,-X 2 -L-T-X 5 -X 6
  • AI-X I -X 2 -V-A-X 5 -X 6 -X 7 -(S-TAMRA)J A ⁇ -X 1 -X 2 -V-A-X 5 -X 6 -X 7 -(S-TAMRA);
  • A5-X I - X 2 -V-A-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X 1 -X 2 -V-A-X 5 -X 6 -X 7 -(S-TAMRA);
  • AI-X I -X 2 -V-Y-X 5 -X 6 -X 7 -(S-TAMRA)J AI-X I -X 2 -V-Y-X 5 -X 6 -X 7 -(S-TAMRA)J
  • A5-X 1 - X 2 -V-Y-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X 1 -X 2 -V-Y-X 5 -X 6 -X 7 -(S-TAMRA)J
  • AI-X 1 -X 2 -K-G-X 5 -X 6 -X 7 -(S-TAMRA)J AZ-X 1 -X 2 -K-G-X 5 -X 6 -X 7 -(S-TAMRA);
  • A3-X 1 -X 2 -K-G-X 5 -X 6 -X 7 -(5-TAMRA)J A4-X I -X 2 -K-G-X 5 -X 6 -X 7 -(5-TAMRA);
  • A5-X I - X 2 -K-G-X 5 -X 6 -X 7 -(S-TAMRA)J AO-X I -X 2 -K-G-X 5 -X 6 -X 7 -(S-TAMRA)J

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Abstract

The present invention is generally directed to biological assays. More specifically it is directed to optimal FRET pairs and methods related to their use. In a composition aspect, the present invention is directed to FRET-based protease substrates selected from the list of substrates shown in the Detailed Description. In a method aspect, the present invention is directed to a method of performing a FRET-based assay, where the assay includes the following steps: adding a solution or suspension containing one or more different proteins to a reaction vessel; adding a liquid comprising a FRET-substrate to the reaction vessel, wherein the FRET substrate is selected from the list of substrates shown in the Detailed Description. In a kit aspect, the present invention is directed to a kit for performing a FRET-based assay, wherein the kit comprises a protease substrate. The protease substrate is selected from the list of substrates shown in the Detailed Description.

Description

OPTIMAL FRET PAIRS AND RELATED METHODS
Field of the Invention
The present invention is generally directed to biological assays. More specifically it is directed to optimal FRET pairs and methods related to their use.
Background of the Invention
Proteases are enzymes that cleave protein substrates to form smaller proteins. The smaller proteins exhibit specific beneficial or detrimental functions in the host organism. Where detrimental functions are exhibited, the proteins oftentimes are the root cause of various diseases. Examples of such diseases include cancer, arthritis, Alzheimer's, hypertension, hepatitis and AIDS.
Pharmaceutical research directed to proteases came to the scientific forefront when researchers realized that an HIV protease could be the key to treating early-stage HIV infections. When the research uncovered several small molecule inhibitors of HIV protease, and the inhibitors proved effective in a clinical setting, discovering new proteases, understanding their function, and using them in the development of novel pharmaceuticals became an industry priority.
The development of protease-based pharmaceuticals required the emergence of new assays that one could use to monitor the effect of small or large molecules on protease activity. Preferred assays were homogeneous, meaning one could obtain a continuous reading of activity rather than stopping enzymatic activity, separating reaction components and then obtaining an activity reading. Useful assays further had to be very sensitive so one could measure activity even when only a small amount of protease was present in the reaction medium. Given the pharmaceutical companies' assay requirements, spectrophotometric- based assays appeared to be the likely answer for probing protease activity; they are homogeneous, fast, accurate and easy to use. Spectrophotometric assays are either fluorogenic or chromogenic, with fluorogenic assays being superior in many cases. This is because the fluorogenic assays have a wider linear dynamic range and offer better reproduceability. Forster resonance energy transfer ("FRET") assays are a type of fluorogenic spectrophotometric assay pharmaceutical companies have oftentimes used for rapid activity screening.
FRET involves the transfer of excited-state energy from one type of compound ("donor") to another type ("acceptor"). The donor typically transfers (i.e., emits) short wavelengths of light that are within the wavelength range the acceptor can absorb. Energy transfer occurs within a specified distance between the donor and acceptor, usually 10 to 100 A. The distance at which energy transfer equals 50 percent is called the Forster radius. Within this distance, a decrease in donor fluorescence can be detected upon donor to acceptor energy transfer.
The use of FRET in a protease assay involves the construction of a protease substrate that includes both a donor and an acceptor positioned on opposite sides of the cleavage site at an appropriate distance from one another. The substrate is typically constructed from a peptide sequence derived from a known protein cleaved by the protease. Spectrophotometric methods allow one to observe whether the protease substrate has been cleaved, since cleavage provides that the donor-acceptance distance exceeds the Forster radius; this results in recovery of the donor's fluorescence. Reports that discuss the use of FRET-based assays for examination of protease activity include, for example: U.S. Pat. No. 6,291,201 assigned to Zeneca Ltd., entitlted "Fluorescence Energy Transfer Substrates"; U.S. Pat. No. 6,445,268 assigned to Iconix Pharmaceuticals Ltd., entitled, "Hydrolytic Enzyme Substrates and Assay Method"; U.S. Pat. No. 7,157,553 assigned to the United States, entitled "Highthroughput Assays for the Proteolytic Activities of Clostridial Neurotoxins". Each of the preceding patents is incorporated-by-reference for all purposes into this document.
Summary of the Invention
The present invention is generally directed to biological assays. More specifically it is directed to optimal FRET pairs and methods related to their use.
In a composition aspect, the present invention is directed to FRET-based protease substrates selected from the list of substrates shown in the Detailed Description above.
In a method aspect, the present invention is directed to a method of performing a FRET-based assay, where the assay includes the following steps: adding a solution or suspension containing one or more different proteins to a reaction vessel; adding a liquid comprising a FRET-substrate to the reaction vessel, wherein the FRET substrate is selected from the list of substrates shown in the Detailed Description section below.
In a kit aspect, the present invention is directed to a kit for performing a FRET- based assay, wherein the kit comprises a protease substrate. The protease substrate is selected from the list of substrates shown in the Detailed Description section below.
Brief Description of the Drawings
FIGS 1-6 show the structures of various members of optimal FRET pairs according to the present invention. Detailed Description of the Invention
With extensive experience, the inventors have developed a set of guidelines that aid in the development of a FRET-based protease substrate. A donor/acceptor pair for inclusion within the substrate should be chosen such that the absorption spectrum of the acceptor overlaps with the emission spectrum of the donor. Generally, a fluorescent donor and a non-fluorescent acceptor are used to make protease peptide substrates. One should keep in mind, however, that the donor/acceptor pair choice may be limited by the fiuorometer filter at hand.
The donor and acceptor molecules must be in close proximity (e.g., 10 to 100 A) in order for the acceptor to effectively quench donor fluorescence. Once an active protease recognizes and cleaves the substrate into two separate fragments, the increase in the donor-acceptor distance causes FRET efficiency to decrease, resulting in the recovery of the donor's fluorescence. The time-dependent increase in fluorescence intensity is related to the extent of substrate hydrolysis.
One may use peptide sequences containing unnatural amino acids or modified bonds to construct the FRET-based substrate, or one may simply use the native peptide containing a protease cleavage site. Use of non-native sequences may be used to increase cleavage efficiency, to protect the peptide from degradation or to increase peptide solubility. For example, an ester linkage can replace an amide linkage in the peptide to increase cleavage efficiency.
Most fluorophores react with amino groups, meaning they can be conjugated to the α-amino group of Lysine. Thiol reactive dyes can be used to conjugate donors and/or acceptors to Cys-containing peptides. This is an economical way to utilize the donors and/or acceptors since the peptides can be HPLC purified first before reacting with the dyes.
For hydrophobic sequences, Lysines or Arginines may be added to increase solubility. These amino acids must be added at the appropriate positions without adversely affecting the protease recognition site.
Using the guidelines above, the inventors discovered a variety of optimal peptide donor-acceptor combinations.
A generic representation of the peptide substrates to which the donor and acceptor are conjugated is shown below, where the FRET moieties area attached at any suitable point along the substrate:
X1-X2-X3-X4-X5-XO-XT-XS-X^XIO-XI I-XU-XB-XM-XIS-XIO-XΠ
and the moieties Xi-X17 are defined as follows:
Xi = D, γ-Abu, K, E, P, G, E, H, A, T, R, F;
X2 = E, S, G, V, H, L5 K, P, Q, Y, Cha, A;
X3 = D, Q, Cha, N, P, A, G, S5 L5 V, K;
X4 = E, N5 Abu, L5 F5 Q5 15 R, G5 A, K, T5 P, Nva, C(Me)5 Y;
X5 = Abu-y-[COO]5 Y5 Smc, D5 H, A5 L, V, P5 G5 S, E5 R5 Dap, W5 K;
X6 = A5 P5 HAr5 L5 V5 F, Q, R5 S5 G, E, M5 H5 Ar5 Dab, or no amino acid;
X7 = S5 15 K, E, V, R, F, A, P, G, H, Nva, Dap, or no amino acid;
X8 = C, V, A, F5 15 S, R5 E, K, P, S, W, Dab, Q5 or no amino acid;
X9 = Q5 K5 H5 S5 L5 A5 1, K(Ac), D, R, or no amino acid; X10 = K, S, A, I, Y, P, L, G, or no amino acid;
X1 I = R, K, A, NIe, L, or no amino acid;
Xi2 = K, F, or no amino acid;
Xi3 = S, or no amino acid;
Xi4 = T, or no amino acid;
X15 = E, or no amino acid;
Xi6 = G, or no amino acid;
Xi 7 = K, or no amino acid.
Nonlimiting examples of peptide substrates to which the donor and acceptor are conjugated include ("*" designating point of FRET moiety attachment):
HCV NS3/4A protease substrate: *-Asp-Glu-Asp-Glu-Glu-Abu-y-[COO]Ala- Ser-Cys(*)-NH2 (SEQ ID NO. 1).
HIV-I protease substrate: *-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(*)-OH (SEQ ID NO. 2).
MMP generic substrate: *-KGP-Cha-Abu-Smc-HAr-K(*)-AK-NH2 (SEQ ID NO. 3).
B-Secretase substrate: *-EVNLDAEFK-* (SEQ ID NO. 4).
Renin substrate: ♦-KHPFHLVIHK-* (SEQ ID NO. 5).
B-secretase (TACE) substrate: *-PLAQ AVRSSSRK-* (SEQ ID NO. 6).
Cathepsin D substrate: ♦-GKPILFFRLK-* (SEQ ID NO. 7).
Cathepsin S substrate: * -EKARVL AE AAK-* (SEQ ID NO. 8).
Cathepsin K substrate: *-HPGGPQK-* (SEQ ID NO. 9).
Aggrecanase-1 substrate: *-AELQGRPISIAK-* (SEQ ID NO. 10). Aggrecanase-2 substrate: * -TESESRGAI YK-* (SEQ ID NO. 11).
Calpain substrate A: ♦-PLLAERPK-* (SEQ ID NO. 12).
Calpain substrate B: ♦-TPLKSPPPSPRK-* (SEQ ID NO. 13).
Sirtuin substrate: *-GQSTSSHS-K(Ac)-L-Nle-FSTEG-K-* (SEQ ID NO. 14).
Thrombin substrate: *-D YVPRGSKDK-* (SEQ ID NO. 15).
MMP-I substrate: *-PLA-Nva-Dap-* (SEQ ID NO. 16).
MMP-2 substrate: * -PLGC(Me)HARK-* (SEQ ID NO. 17).
MMP-3 substrate: * -RPKP VE-Nva- WRK-* (SEQ ID NO. 18).
MMP-7 substrate: *-P YAYWMRK-* (SEQ ID NO. 19).
MMP-8 substrate: *-P-Cha-G-Nva-HA-Dap-* (SEQ ID NO. 20).
MMP-9 substrate: *-γ-Abu-P-Cha-Abu-Smc-HA-Dab-* (SEQ ID NO. 21).
MMP-IO substrate: * -RPKPL A-Nva- WK-* (SEQ ID NO. 22).
MMP- 12 substrate: *-PLGLEEAK-* (SEQ ID NO. 23).
MMP-13 substrate: *-PLGLWArK-* (SEQ ID NO. 24).
MMP- 14 substrate: *-γ-Abu-PQGL-Dab-* (SEQ ID NO. 25).
West Nile Virus Protease substrate: *-FASGKRSQIGLK-* (SEQ ID NO. 26).
FRET substrates - i.e., peptide/donor/acceptor combinations - according to the present invention are shown below. "Al" through "A6", "Bl" through "B6", "Cl" through "C4" and 5-TAMRA designate moieties of a FRET pair. (See FIGS 1-6.) The moiety is attached through a reaction with an activated carboxylic acid group of an α,β- unsaturated sulfone group, with the point of attachment shown as "*". "5-TAMRA" is a rhodamine dye (i.e., 5-carboxytetramethylrhodamine) and is attached to the substrate using chemistry known to those of skill in the art. Nonlimiting examples of optimal FRET pairs/substrate combinations include:
C 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B 1 )-OH; C 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B2)-OH; Cl-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; Cl-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; Cl-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; Cl-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(B6)-OH;
C2-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B 1 )-OH; C2-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B2)-OH; C2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C2-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; C2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(B6)-OH;
C3 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B 1 )-OH; C3 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-πe-Val-Gln-Lys(B2)-OH; C3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C3-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile- VaI-GIn-LyS(BS)-OHj CS-Y-AbU-SCr-GIn-ASn-TyF-PrO-IIe- VaI- Gln-Lys(B6)-OH;
C4-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-LysCB 1 )-OH; C4-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B2)-OH; C4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C4-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; C4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(B6)-OH; B 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C 1 )-OH; B 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B 1 -γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln- Lys(C3)-OH; B 1 -γ- Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH;
B2-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B2-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH;
B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(C I)-OH; B3-γ-Abu-Ser-Gln- Asn-Tyr-Pro-De-Val-Gln-Lys(C2)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH;
B4-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B3 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH;
B5-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B5-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH;
B6-γ- Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B6-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B6-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B6-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH;
Nonlimiting examples of substrates including Al through A6 and 5-TAMRA FRET pairs include:
A 1 -KGP-Cha- Abu-Smc-HAr-K(5-TAMRA)-AK-NH2; A2-KGP-Cha-Abu-Smc- HAr-K(5-TAMRA)-AK-NH2; A3-KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)-AK-NH2; A4-KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)-AK-NH2; A5-KGP-Cha-Abu-Smc-HAr- K(5-TAMRA)-AK-NH2; A6-KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)- AK-NH2;
Al -EVNLDAEFK-(S-TAMRA); A2-EVNLDAEFK-(5 -TAMRA); A3- EVNLDAEFK-(5-TAMRA); A4-EVNLDAEFK-(5-TAMRA); A5-EVNLDAEFK-(5- TAMRA); A6-EVNLDAEFK-(5-TAMRA);
AI-KHPFHLVIHK-(S-TAMRA); A2-KHPFHLVIHK-(5-TAMRA); A3- KHPFHLVIHK-(5-TAMRA); A4-KHPFHLVIHK-(5 -TAMRA); A5 -KHPFHL VIHK-(5- TAMRA); A6-KHPFHLVIHK-(5-TAMRA);
Al-PLAQAVRSSSRK-(5-TAMRA); A2-PLAQAVRSSSRK-(5-TAMRA); A3- PLAQAVRSSSRK-(5-TAMRA); A4-PLAQAVRSSSRK-(5-TAMRA); A5- PLAQAVRSSSRK-(5-TAMRA); A6-PLAQAVRSSSRK-(5-TAMRA);
Al-GKPILFFRLK-(5-TAMRA); A2-GKPILFFRLK-(5-TAMRA); A3- GKPILFFRLK-(5-TAMRA); A4-GKPILFFRLK-(5-TAMRA); A5-GKPILFFRLK-(5- TAMRA); A6-GKPILFFRLK-(5-TAMRA);
Al-EKARVLAEAAK-(5-TAMRA); A2-EKARVLAEAAK-(5-TAMRA); A3- EKARVLAEAAK-(5-TAMRA); A4-EKARVLAEAAK-(5-TAMRA); A5- EKARVLAEAAK-(5-TAMRA); A6-EKARVLAEAAK-(5-TAMRA);
Al-HPGGPQK-(5-TAMRA); A2-HPGGPQK-(5-TAMRA); A3-HPGGPQK-(5- TAMRA); A4-HPGGPQK-(5-TAMRA); A5-HPGGPQK-(5-TAMRA); A6-HPGGPQK- (5-TAMRA);
Al-AELQGRPISIAK-(5-TAMRA); A2-AELQGRPISIAK-(5 -TAMRA); A3- AELQGRPISIAK-(5-TAMRA); A4-AELQGRPISIAK-(5 -TAMRA); A5- AELQGRPISIAK-(5-TAMRA); A6-AELQGRPISIAK-(5-TAMRA); AI-TESESRGAIYK-(S-TAMRA); AZ-TESESRGAIYK-(S-TAMRA); A3- TESESRGAIYK-(5-TAMRA); A4-TESESRGAIYK-(5-TAMRA); A5-TESESRGAIYK- (5-TAMRA); A6-TESESRGAIYK-(5 -TAMRA);
Al-PLLAERPK-(5-TAMRA); A2-PLLAERPK-(5-TAMRA); A3-PLLAERPK- (5-TAMRA); A4-PLLAERPK-(5-TAMRA); A5-PLLAERPK-(5-TAMRA); A6- PLLAERPK-(5-TAMRA);
Al-TPLKSPPPSPRK-(5-TAMRA); A2-TPLKSPPPSPRK-(5-TAMRA); A3- TPLKSPPPSPRK-(5-TAMRA); A4-TPLKSPPPSPRK-(5-TAMRA); A5- TPLKSPPPSPRK-(S-TAMRA)J AO-TPLKSPPPSPRK-(S-TAMRA);
Al-GQSTSSHS-K(Ac)-L-Nle-FSTEG-K-(5-TAMRA); A2-GQSTSSHS-K(Ac)- L-NIe-FSTEG-K-(S-TAMRA)J AS-GQSTSSHS-K(Ac)-L-NIe-FSTEG-K-(S-TAMRA); A4-GQSTSSHS-K(Ac)-L-Nle-FSTEG-K-(5-TAMRA); A5-GQSTSSHS-K(Ac)-L-Nle- FSTEG-K-(S-TAMRA)J AO-GQSTSSHS-K(Ac)-L-NIe-FSTEG-K-(S-TAMRA);
Al-DYVPRGSKDK-(5-TAMRA); A2-DYVPRGSKDK-(5-TAMRA); A3- DYVPRGSKDK-(5-TAMRA); A4-DYVPRGSKDK-(5 -TAMRA); A5 -D YVPRGSKDK- (5-TAMRA); A6-DYVPRGSKDK-(5-TAMRA);
Al-PLA-Nva-Dap-(5-TAMRA); A2-PLA-Nva-Dap-(5-TAMRA); A3-PLA-Nva- Dap-(5-TAMRA); A4-PLA-Nva-Dap-(5-TAMRA); A5-PLA-Nva-Dap-(5-TAMRA); A6-PLA-Nva-Dap-(5-TAMRA);
Al-PLGC(Me)HARK-(5-TAMRA); A2-PLGC(Me)HARK-(5-TAMRA); A3- PLGC(Me)HARK-(5-TAMRA); A4-PLGC(Me)HARK-(5-TAMRA); A5- PLGC(Me)HARK-(5-TAMRA); A6-PLGC(Me)HARK-(5-TAMRA); A 1 -RPKPVE-NVa-WRK-(S-TAMRA); A2-RPKPVE-Nva- WRK-(5-TAMRA); A3-RPKPVE-Nva-WRK-(5-TAMRA); A4-RPKPVE-Nva-WRK-(5-TAMRA); A5- RPKPVE-Nva-WRK-(5-TAMRA); A6-RPKPVE-Nva-WRK-(5-TAMRA);
A 1-PYAYWMRK-(S-TAMRA); A2-PYAYWMRK-(5-TAMRA); A3- PYAYWMRK-(5-TAMRA); A4-PYAYWMRK-(5-TAMRA); A5-PYAYWMRK-(5- TAMRA); A6-PYAYWMRK-(5-TAMRA);
Al -P-Cha-G-Nva-HA-Dap-(5-TAMRA); A2-P-Cha-G-Nva-HA-Dap-(5- TAMRA); A3-P-Cha-G-Nva-HA-Dap-(5-TAMRA); A4-P-Cha-G-Nva-HA-Dap-(5- TAMRA); A5-P-Cha-G-Nva-HA-Dap-(5-TAMRA); A6-P-Cha-G-Nva-HA-Dap-(5- TAMRA);
Al -γ-Abu-P-Cha-Abu-Smc-HA-Dab-(5-TAMRA); A2-γ-Abu-P-Cha-Abu-Smc- HA-Dab-(5-TAMRA); A3-γ-Abu-P-Cha-Abu-Smc-HA-Dab-(5-TAMRA); A4-γ-Abu-P- Cha-Abu-Smc-HA-Dab-(5-TAMRA); A5-γ-Abu-P-Cha-Abu-Smc-HA-Dab-(5-TAMRA); A6-γ-Abu-P-Cha-Abu-Smc-HA-Dab-(5-TAMRA);
Al-RPKPLA-Nva-WK-(5-TAMRA); A2-RPKPLA-Nva-WK-(5-TAMRA); A3- RPKPLA-Nva-WK-(5-T AMRA); A4-RPKPLA-Nva-WK-(5-TAMRA); A5-RPKPLA- Nva-WK-(5-TAMRA); A6-RPKPLA-Nva-WK-(5-TAMRA);
Al-PLGLEEAK-(5-TAMRA); A2-PLGLEEAK-(5 -TAMRA); A3-PLGLEEAK- (5-TAMRA); A4-PLGLEEAK-(5 -TAMRA); A5 -PLGLEE AK-(5 -TAMRA); A6- PLGLEEAK-(5-TAMRA);
Al-PLGLWArK-(5-TAMRA); A2-PLGLWAΓK-(5-TAMRA); A3-PLGL WArK- (5-TAMRA); A4-PLGLWArK-(5-TAMRA); A5-PLGLWArK-(5-TAMRA); A6- PLGLW ArK-(5-TAMRA); Al-γ-Abu-PQGL-Dab-(5-TAMRA); A2-γ-Abu-PQGL-Dab-(5-TAMRA); A3-γ- Abu-PQGL-Dab-(5-TAMRA); A4-γ-Abu-PQGL-Dab-(5-TAMRA); A5-γ-Abu-PQGL- Dab-(5-TAMRA); A6-γ-Abu-PQGL-Dab-(5-TAMRA);
AI -FASGKRSQIGLK-(S-TAMRA); AZ-FASGKRSQIGLK-(S-TAAdRA); A3- FASGKRSQIGLK-(5-TAMRA); A4-FASGKRSQIGLK-(5-TAMRA); A5- FASGKRSQIGLK-(5-TAMRA); A6-FASGKRSQIGLK-(5-TAMRA).
The same substrates (e.g., * -EVNLDAEFK-*) above may include optimal FRET pairs selected from Bl through B6 combined with Cl through C4 in a manner similar to the nonlimiting examples of substrates including optimal Al through A6 and 5 -TAMRA FRET pairs shown above.
Nonlimiting examples of generic substrates including Al through A6 and 5- TAMRA FRET pairs include (where moieties are defined as above):
AI-X1-X2-X3-X4-X5-X6-X7-XS-X^XIO-XI I-XI2-XI3-XH-XI 5-XI6-XI7-(S-TAMRA); A2-Xi-X2-X3-X4-X5-X6-X7-X8-X9-Xio-Xπ-Xi2-Xi3-Xi4-Xi5-Xi6-Xi7-(5-TAMRA); A3- XI-X2-X3-X4-X5-X6-X7-X8-X^XIO-XiI-Xn-Xi3-XM-Xi5-Xi6-Xi7-(S-TAMRA); A4-X,- X2-X3-X4-X5-X6-X7-X8-X9-XIO-XI I-XI2-XI3-XI4-XI 5-XI6-XI7-(S-TAMRA); A5-X,-X2- X3-X4-X5-X6-X7-X8-X9-XIO-XI I-XI2-XI3-XI4-XI5-XI6-XI7-(S-TAMRA)J AO-X1-X2-X3- X4-X5-X6-X7-X8-X9-XIO-XU-XI2-XI3-XI4-XI 5-XI6-XI7-(S-TAMRA);
Al-X1-X2-D-X4-X5-X6-X7-(S-TAMRA); A2-XI-X2-D-X4-X5-X6-X7-(5-TAMRA); AS-Xi-X2-D-X4-X5-X6-X7-(S-TAMRA); A4-X,-X2-D-X4-X5-X6-X7-(5-TAMRA); A5- X1-X2-D-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-X4-X5-X6-X7-(S-TAMRA); Al-Xi-X2-D-E-X5-X6-X7-(S-TAMRA); A2-X,-X2-D-E-X5-X6-X7-(5-TAMRA); A3-X,-X2-D-E-X5-X6-X7-(5-TAMRA); A4-Xi-X2-D-E-X5-X6-X7-(5-TAMRA); A5-Xi- X2-D-E-X5-X6-X7-(S-TAMRA)J AO-XI-X2-D-E-X5-X6-X7-(S-TAMRA);
Al-X1-X2-D-N-X5-X6-X7-(S-TAMRA); A2-X,-X2-D-N-X5-X6-X7-(5-TAMRA); A3-Xi-X2-D-N-X5-X6-X7-(5-TAMRA); A4-X,-X2-D-N-X5-X6-X7-(5-TAMRA); A5-X,- X2-D-N-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-N-X5-X6-X7-(S-TAMRA);
Al-X1-X2-D-AbU-X5-X6-X7-(S-TAMRA); A2-X, -X2-D- Abu-X5-X6-X7-(5- TAMRA); AS-X1-X2-D-AbU-X5-X6-X7-(S-TAMRA); A4-X!-X2-D-Abu-X5-X6-X7-(5- TAMRA); AS-X1 -X2-D- Abu-X5-X6-X7-(5-TAMRA); A6-X1 -X2-D- Abu-X5-X6-X7-(5- TAMRA);
Al-X1-X2-D-L-X5-X6-X7-(S-TAMRA); A2-X!-X2-D-L-X5-X6-X7-(5-TAMRA); AS-X1-X2-D-L-X5-X6-X7-(S-TAMRA); A4-Xi-X2-D-L-X5-X6-X7-(5-TAMRA); A5-Xi- X2-D-L-X5-X6-X7-(5-TAMRA); AO-Xi-X2-D-L-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-D-F-X5-X6-X7-(5-TAMRA); A2-Xi-X2-D-F-X5-X6-X7-(5-TAMRA); A3-X1-X2-D-F-X5-X6-X7-(5-TAMRA); A4-Xi-X2-D-F-X5-X6-X7-(5-TAMRA); A5-Xi- X2-D-F-X5-X6-X7-(S-TAMRA)J AO-XI-X2-D-F-X5-X6-X7-(S-TAMRA);
AI-X1-X2-D-Q-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-D-Q-X5-X6-X7-(S-TAMRA); AS-X1-X2-D-Q-X5-X6-X7-(S-TAMRA); A4-X,-X2-D-Q-X5-X6-X7-(5-TAMRA); A5-XI- X2-D-Q-X5-X6-X7-(S-TAMRA)J AO-XI-X2-D-Q-X5-X6-X7-(S-TAMRA);
AI-XI-X2-D-I-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-D-I-X5-X6-X7-(S-TAMRA); AS-X1-X2-D-I-X5-X6-X7-(S-TAMRA); A4-X,-X2-D-I-X5-X6-X7-(5-TAMRA); A5-XI- X2-D-I-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-I-X5-X6-X7-(S-TAMRA); Al -Xi-X2-D-R-X5-X6-X7-(S-TAMRA); A2-X( -X2-D-R-X5-X6-X7-(S -TAMRA); A3-X,-X2-D-R-X5-X6-X7-(5-TAMRA); A4-X,-X2-D-R-X5-X6-X7-(5-TAMRA); A5-X,- X2-D-R-X5-X6-X7-(S-TAMRA); A6-Xi-X2-D-R-X5-X6-X7-(5-TAMRA);
Al-X1-X2-D-G-X5-X6-X7-(S-TAMRA); A2-Xi-X2-D-G-X5-X6-X7-(5-TAMRA); AS-X1-X2-D-G-X5-X6-X7-(S-TAMRA); A4-X1-X2-D-G-X5O£6-X7-(5-TAMRA); A5-Xr X2-D-G-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-G-X5-X6-X7-(S-TAMRA);
Al-X1-X2-D-A-X5-X6-X7-(S-TAMRA); A2-X1-X2-D-A-X5-X6-X7-(5-TAMRA); AS-XI-X2-D-A-X5-X6-X7-(S-TAMRA)J A^XI-X2-D-A-X5-X6-X7-(S-TAMRA)J AS-X1- X2-D-A-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-A-X5-X6-X7-(S-TAMRA);
Al-X1-X2-D-K-X5-X6-X7-(S-TAMRA); A2-X,-X2-D-K-X5-X6-X7-(5-TAMRA); AS-X1-X2-D-K-X5-X6-X7-(S-TAMRA); A4-X,-X2-D-K-X5-X6-X7-(5-TAMRA); A5-X!- X2-D-K-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-K-X5-X6-X7-(S-TAMRA)J
Al-X1-X2-D-T-X5-X6-X7-(S-TAMRA); A2-X1-X2-D-T-X5-X6-X7-(5-TAMRA); AS-X1-X2-D-T-X5-X6-X7-(S-TAMRA); A4-X,-X2-D-T-X5-X6-X7-(5-TAMRA); A5-Xi- X2-D-T-X5-X6-X7-(5-TAMRA); A6-X, -X2-D-T-X5-X6-X7-(5 -TAMRA);
Al-X1-X2-D-P-X5-X6-X7-(S-TAMRA); A2-X,-X2-D-P-X5-X6-X7-(5-TAMRA); A3-Xi -X2-D-P-X5-X6-X7-(S-TAMRA); A4-X, -X2-D-P-X5-X6-X7-(S-TAMRA); A5-X, - X2-D-P-X5-X6-X7-(S-TAMRA)J AO-X1-X2-D-P-X5-X6-X7-(S-TAMRA);
Al-X1-X2-D-NVa-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-D-NVa-X5-X6-X7-(S- TAMRA)J AS-X1-X2-D-NVa-X5-X6-X7-(S-TAMRA); A4-Xi-X2-D-Nva-X5-X6-X7-(5- TAMRA)J AS-X1-X2-D-NVa-X5-X6-X7-(S-TAMRA)^o-Xi-X2-D-NVa-X5-X6-X7-(S- TAMRA); Al-X1-X2-D-C(Me)-X5-X6-X7-(S-TAMRA); A2-X,-X2-D-C(Me)-X5-X6-X7-(5- TAMRA); A3-X,-X2-D-C(Me)-X5-X6-X7-(5-TAMRA); A4-X,-X2-D-(CMe)-X5-X6-X7- (5-TAMRA); A5-Xi-X2-D-C(Me)-X5-X6-X7-(5-TAMRA); AO-X1-X2-D-C(Me)-X5-X6- X7-(5-TAMRA);
Al-X1-X2-D-Y-X5-X6-X7-(S-TAMRA); A2-X!-X2-D-Y-X5-X6-X7-(5-TAMRA); AS-X1-X2-D-Y-X5-X6-X7-(S-TAMRA); A4-X1-X2-D-Y-X5-X6-X7-(5-TAMRA); A5-X,- X2-D-Y-X5-X6-X7-(S-TAMRA)^O-X1-X2-D-Y-X5-X6-X7-(S-TAMRA);
AI-X1-X2-Q-X4-X5-X6-X7-(S-TAMRA)J AI-X1-X2-Q-X4-X5-X6-X7-(S-TAMRA); AS-X1-X2-Q-X4-X5-X6-X7-(S-TAMRA); A4-X1-X2-Q-X4-X5-X6-X7-(5-TAMRA); A5- X1-X2-Q-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-Q-X4-X5-X6-XV-(S-TAMRA);
Al-X1-X2-Q-L-X5-X6-X7-(S-TAMRA); A2-X,-X2-Q-L-X5-X6-X7-(5-TAMRA); AS-X1-X2-Q-L-X5-X6-X7-(S-TAMRA); A4-Xi-X2-Q-L-X5-X6-X7-(5-TAMRA); A5-Xi- X2-Q-L-X5-X6-X7-(S-TAMRA); AO-X1-X2-Q-L-X5-X6-X7-(S-TAMRA);
Al-X1-X2-Q-F-X5-X6-X7-(S-TAMRA); A2-X1-X2-Q-F-X5-X6-X7-(5-TAMRA); AS-X1-X2-Q-F-X5-X6-X7-(S-TAMRA); A4-Xi-X2-Q-F-X5-X6-X7-(5-TAMRA); A5-Xr X2-Q-F-X5-X6-X7-(S-TAMRA)^O-X1-X2-Q-F-X5-X6-X7-(S-TAMRA);
AI-XI-X2-Q-Q-X5-X6-X7-(S-TAMRA)J AI-XI-X2-Q-Q-X5-X6-X7-(S-TAMRA); A3-Xi-X2-Q-Q-X5-X6-X7-(5-TAMRA); A4-X1-X2-Q-Q-X5-X6-X7-(5-TAMRA); A5-Xi- X2-Q-Q-X5-X6-X7-(S-TAMRA)J AO-X1-X2-Q-Q-X5-X6-X7-(S-TAMRA);
Al-X,-X2-Q-I-X5-X6-X7-(5-TAMRA); A2-X,-X2-Q-I-X5-X6-X7-(5-TAMRA); A3-Xi -X2-Q-I-X5-X6-X7-(5-TAMRA); A4-X, -X2-Q-I-X5-X6-X7-(S-TAMRA); A5-X] - X2-Q-I-X5-X6-X7-(S-TAMRA)J AO-XI-X2-Q-I-X5-X6-X7-(S-TAMRA); AI-X1-X2-Q-R-X5-X6-X7-(S-TAMRA)S AI-X1-X2-Q-R-X5-X6-X7-(S-TAMRA); A3-X,-X2-Q-R-X5-X6-X7-(5-TAMRA); A4-X,-X2-Q-R-X5-X6-X7-(5-TAMRA); A5-Xr X2-Q-R-X5-X6-X7-(S-TAMRA)J AO-X1-X2-Q-R-X5-X6-X7-(S-TAMRA);
A1-X,-X2-Q-G-X5-X6-X7-(5-TAMRA); A2-XI-X2-Q-G-X5-X6-X7-(5-TAMRA);
A3-X1-X2-Q-G-X5-X6-X7-(5-TAMRA); A4-X1-X2-Q-G-X5-X6-X7-(5-TAMRA); A5-Xr X2-Q-G-X5-X6-X7-(S-TAMRA)J AO-XI-X2-Q-G-X5-X6-X7-(S-TAMRA);
Al-X1-X2-Q-A-X5-X6-X7-(S-TAMRA); A2-Xj-X2-Q-A-X5-X6-X7-(5-TAMRA); AS-X1-X2-Q-A-X5-X6-X7-(S-TAMRA); A4-X, -X2-Q- A-X5-X6-X7-(S-TAMRA); A5-Xr X2-Q-A-X5-X6-X7-(S-TAMRA); Ao-Xi-X2-Q-A-X5-X6-X7-(S-TAMRA);
AI-X1-X2-Q-K-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-Q-K-X5-X6-X7-(S-TAMRA); A3-XI-X2-Q-K-X5-X6-X7-(5-TAMRA); A4-X,-X2-Q-K-X5-X6-X7-(5-TAMRA); A5-X1- X2-Q-K-X5-X6-X7-(S-TAMRA)J AO-XI-X2-Q-K-X5-X6-X7-(S-TAMRA);
Al-X1-X2-Q-T-X5-X6-X7-(5-TAMRA); A2-X,-X2-Q-T-X5-X6-X7-(5-TAMRA); A3-Xi-X2-Q-T-X5-X6-X7-(5-TAMRA); A4-X,-X2-Q-T-X5-X6-X7-(5-TAMRA); A5-Xi- X2-Q-T-X5-X6-X7-(5-TAMRA); A6-X1 -X2-Q-T-X5-X6-X7-(5-TAMRA);
A 1 -Xi -X2-Q-P-X5-X6-X7-(S-TAMRA); A2-X, -X2-Q-P-X5-X6-X7-(S-TAMRA); A3-Xi-X2-Q-P-X5-X6-X7-(5-TAMRA); A4-Xi-X2-Q-P-X5-X6-X7-(5-TAMRA); A5-X]- X2-Q-P-X5-X6-X7-(S-TAMRA)J AO-X1-X2-Q-P-X5-X6-X7-(S-TAMRA);
Al-X1-X2-Q-NVa-X5-X6-X7-(S-TAMRA); A2-X,-X2-Q-Nva-X5-X6-X7-(5- TAMRA); A3-Xi-X2-Q-Nva-X5-X6-X7-(5-TAMRA); A4-Xi-X2-Q-Nva-X5-X6-X7-(5- TAMRA); A5-X,-X2-Q-Nva-X5-X6-X7-(5-TAMRA); AO-X1-X2-Q-NVa-X5-X6-X7-(S- TAMRA); Al-X,-X2-Q-C(Me)-X5-X6-X7-(5-TAMRA); A2-Xi-X2-Q-C(Me)-X5-X6-X7-(5- TAMRA); AS-X1-X2-Q-C(Me)-X5-X6-X7-(S-TAMRA); A4-X, -X2-Q-(CMe)-X5-X6-X7- (5-TAMRA); AS-X1-X2-Q-C(Me)-X5-X6-X7-(S-TAMRA)^O-X1-X2-Q-C(Me)-X5-X6- X7-(5-TAMRA);
AI-X1-X2-Q-Y-X5-X6-X7-(S-TAMRA)J AI-X1-X2-Q-Y-X5-X6-X7-(S-TAMRA); A3-X,-X2-Q-Y-X5-X6-X7-(5-TAMRA); A4-X1-X2-Q-Y-X5-X6-X7-(5-TAMRA); A5-X1- X2-Q-Y-X5-X6-X7-(S-TAMRA)J AO-X1-X2-Q-Y-X5-X6-X7-(S-TAMRA);
Al-X1-X2-ClIa-X4-X5-X6-X7-(S-TAMRA); A2-Xi-X2-Cha-X4-X5-X6-X7-(5- TAMRA); AS-X1-X2-ClIa-X4-X5-X6-X7-(S-TAMRA); A^Xi-X2-ClIa-X4-X5-X6-X7-(S- TAMRA)J AS-X1-X2-ClIa-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-ClIa-X4-X5-X6-X7-(S- TAMRA);
Al-Xi-X2-Cha-L-X5-X6-X7-(5-TAMRA); A2-Xi-X2-Cha-L-X5-X6-X7-(5- TAMRA)J AS-X1-X2-ClIa-L-X5-X6-X7-(S-TAMRA); A4-X1-X2-Cha-L-X5-X6-X7-(5- TAMRA)J AS-X1-X2-CHa-L-X5-X6-X7-(S-TAMRA)J Ao-X1-X2-ClIa-L-X5-X6-X7-(S- TAMRA);
Al-X1-X2-ClIa-F-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-ClIa-F-X5-X6-X7-(S- TAMRA)J A3-Xi-X2-Cha-F-X5-X6-X7-(5-TAMRA); A4-X!-X2-Cha-F-X5-X6-X7-(5- TAMRA); AS-X1-X2-CHa-F-X5-X6-X7-(S-TAMRA)J AO-X1-X2-ClIa-F-X5-X6-X7-(S- TAMRA);
A 1 -X1 -X2-ClIa-Q-X5-X6-X7-(S-TAMRA); A2-X,-X2-Cha-Q-X5-X6-X7-(5- TAMRA); A3-X, -X2-CIm-Q-X5-X6-X7-(S-TAMRA); A4-X!-X2-Cha-Q-X5-X6-X7-(5- TAMRA)J AS-X1-X2-CHa-Q-X5-X6-X7-(S-TAMRA)J AO-X1-X2-ClIa-Q-X5-X6-X7-(S- TAMRA); Al-X,-X2-Cha-I-X5-X6-X7-(5-TAMRA); A2-X,-X2-Cha-I-X5-X6-X7-(5-TAMRA); A3-Xi-X2-Cha-I-X5-X6-X7-(5-TAMRA); A4-Xi-X2-Cha-I-X5-X6-X7-(5-TAMRA); A5- X1-X2-CHa-I-X5-X6-X7-(S-TAMRA)J Ao-Xi-X2-ClIa-I-X5-X6-X7-(S-TAMRA);
Al-X,-X2-Cha-R-X5-X6-X7-(5-TAMRA); A2-X,-X2-Cha-R-X5-X6-X7-(5- TAMRA); A3-Xi-X2-Cha-R-X5-X6-X7-(5-TAMRA); A4-X,-X2-Cha-R-X5-X6-X7-(5- TAMRA); A5-Xi-X2-Cha-R-X5-X6-X7-(5-TAMRA); A6-Xi-X2-Cha-R-X5-X6-X7-(5- TAMRA);
Al -X1 -X2-Cha-G-X5-X6-X7-(5-TAMRA); A2-X, -X2-Cha-G-X5-X6-X7-(5- TAMRA); A3-XrX2-Cha-G-X5-X6-X7-(5-TAMRA); A4-Xi-X2-Cha-G-X5-X6-X7-(5- TAMRA); A5-Xi-X2-Cha-G-X5-X6-X7-(5-TAMRA); A6-Xi-X2-Cha-G-X5-X6-X7-(5- TAMRA);
Al-X,-X2-Cha-A-X5-X6-X7-(5-TAMRA); A2-Xi-X2-Cha-A-X5-X6-X7-(5- TAMRA); A3-X,-X2-Cha-A-X5-X6-X7-(5-TAMRA); A4-Xi-X2-Cha-A-X5-X6-X7-(5- TAMRA); A5-Xi-X2-Cha-A-X5-X6-X7-(5-TAMRA); A6-X,-X2-Cha-A-X5-X6-X7-(5- TAMRA);
Al-X1-X2-ClIa-K-X5-X6-X7-(S-TAMRA); A2-X,-X2-Cha-K-X5-X6-X7-(5- TAMRA); A3-Xi-X2-Cha-K-X5-X6-X7-(5-TAMRA); A4-X1-X2-Cha-K-X5-X6-X7-(5- TAMRA); A5-X, -X2-Cha-K-X5-X6-X7-(5-TAMRA); A6-X1 -X2-Cha-K-X5-X6-X7-(5- TAMRA);
Al -Xi -X2-Cha-T-X5-X6-X7-(5-TAMRA); A2-X, -X2-ClIa-T-X5-X6-X7-(S- TAMRA); A3-Xj -X2-ClIa-T-X5-X6-X7-(S-TAMRA); A4-X, -X2-ClIa-T-X5-X6-X7-(S- TAMRA); A5-Xi-X2-Cha-T-X5-X6-X7-(5-TAMRA); A6-Xi-X2-Cha-T-X5-X6-X7-(5- TAMRA); A 1 -X1-X2-ClIa-P-X5-X6-X7-(S-TAMRA); A2-X, -X2-Cha-P-X5-X6-X7-(5- TAMRA); A3-Xi-X2-Cha-P-X5-X6-X7-(5-TAMRA); A4-X, -X2-ClIa-P-X5-X6-X7-(S- TAMRA); AS-X1-X2-ClIa-P-X5-X6-X7-(S-TAMRA); A6-X,-X2-Cha-P-X5-X6-X7-(5- TAMRA);
Al-X1-X2-ClIa-NVa-X5-X6-X7-(S-TAMRA); A2-X,-X2-Cha-Nva-X5-X6-X7-(5- TAMRA); AS-X1-X2-ClIa-NVa-X5-X6-X7-(S-TAMRA); A4-X!-X2-Cha-Nva-X5-X6-X7- (5-TAMRA)J AS-X1-X2-ClIa-NVa-X5-X6-X7-(S-TAMRA)J AO-X1-X2-ClIa-NVa-X5-X6- X7-(5-TAMRA);
Al-X1-X2-ClIa-C(Me)-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-ClIa-C(Me)-X5-X6-X7- (5-TAMRA); AS-Xi-X2-ChH-C(Me)-X5-X6-X7-(S-TAMRA); A4-Xi-X2-Cha-(CMe)-X5- X6-X7-(5-TAMRA); AS-X1-X2-CHa-C(Me)-X5-X6-X7-(S-TAMRA); A6-Xi-X2-Cha- C(Me)-X5-X6-X7-(5-TAMRA);
Al-Xi-X2-Cha- Y-X5-X6-X7-(S-TAMRA); A2-Xi-X2-Cha-Y-X5-X6-X7-(5- TAMRA); A3-Xi-X2-Cha-Y-X5-X6-X7-(5-TAMRA); A4-X!-X2-Cha- Y-X5-X6-X7-(5- TAMRA)J AS-X1-X2-CHa-Y-X5-X6-X7-(S-TAMRA)J Ao-Xi-X2-ClIa-Y-X5-X6-X7-(S- TAMRA);
Al-Xi-X2-N-X4-X5-X6-X7-(S-TAMRA); A2-Xi-X2-N-X4-X5-X6-X7-(5-TAMRA); AS-Xi-X2-N-X4-X5-X6-X7-(S-TAMRA); A4-X,-X2-N-X4-X5-X6-X7-(5-TAMRA); A5- X1-X2-N-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-N-X4-X5-X6-X7-(S-TAMRA);
Al -Xi-X2-N-L-X5-X6-X7-(S-TAMRA); A2-X!-X2-N-L-X5-X6-X7-(5-TAMRA); A3-Xi-X2-N-L-X5-X6-X7-(5-TAMRA); A4-X,-X2-N-L-X5-X6-X7-(5-TAMRA); A5-Xj- X2-N-L-X5-X6-X7-(S-TAMRA); A6-X, -X2-N-L-X5-X6-X7-(S-TAMRA); AI-XI-X2-N-F-X5-X6-X7-(S-TAMRA)J AI-X1-X2-N-F-X5-X6-X7-(S-TAMRA); A3-X,-X2-N-F-X5-X6-X7-(5-TAMRA); A4-X1-X2-N-F-X5-X6-X7-(5-TAMRA); A5-XI- X2-N-F-X5-X6-X7-(S-TAMRA)J AO-X1-X2-N-F-X5-X6-X7-(S-TAMRA);
Al-X1-X2-N-Q-X5-X6-X7-(S-TAMRA); A2-X,-X2-N-Q-X5-X6-X7-(5-TAMRA); AS-X1-X2-N-Q-X5-X6-X7-(S-TAMRA); A4-X,-X2-N-Q-X5-X6-X7-(5-TAMRA); A5-X,- X2-N-Q-X5-X6-X7-(S-TAMRA)J AO-XI-X2-N-Q-X5-X6-X7-(S-TAMRA);
Al -Xi-X2-N-I-X5-X6-X7-(S-TAMRA); A2-X,-X2-N-I-X5-X6-X7-(5-TAMRA); AS-X1-X2-N-I-X5-X6-X7-(S-TAMRA); A4-X,-X2-N-I-X5-X6-X7-(5-TAMRA); A5-Xi- X2-N-I-X5-X6-X7-(5-TAMRA); Ao-Xi-X2-N-I-X5-X6-X7-(S-TAMRA);
Al -Xi-X2-N-R-X5-X6-X7-(S-TAMRA); A2-Xi-X2-N-R-X5-X6-X7-(5-TAMRA); A3-Xi-X2-N-R-X5-X6-X7-(5-TAMRA); A4-Xi-X2-N-R-X5-X6-X7-(5-TAMRA); A5-X,- X2-N-R-X5-X6-X7-(S-TAMRA)J AO-XI-X2-N-R-X5-X6-X7-(S-TAMRA);
AI-XI-X2-N-G-X5-X6-X7-(S-TAMRA)J AI-XI-X2-N-G-X5-X6-X7-(S-TAMRA)J A3-X,-X2-N-G-X5-X6-X7-(5-TAMRA); A4-X,-X2-N-G-X5-X6-X7-(5-TAMRA); A5-X1- X2-N-G-X5-X6-X7-(S-TAMRA)J AO-X1-X2-N-G-X5-X6-X7-(S-TAMRA)J
A1-XI-X2-N-A-X5-X6-X7-(5-TAMRA); A2-XI-X2-N-A-X5-X6-X7-(5-TAMRA); AS-Xi-X2-N-A-X5-X6-X7-(S-TAMRA); A4-Xi-X2-N-A-X5-X6-X7-(5-TAMRA); A5-Xi- X2-N-A-X5-X6-X7-(S-TAMRA^ AO-XI-X2-N-A-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-N-K-X5-X6-X7-(S-TAMRA); A2-XI-X2-N-K-X5-X6-X7-(5-TAMRA); AS-Xi-X2-N-K-X5-X6-X7-(S-TAMRA); A4-X,-X2-N-K-X5-X6-X7-(5-TAMRA)j A5-X,- X2-N-K-X5-X6-X7-(S-TAMRA)J AO-XI-X2-N-K-X5-X6-X7-(S-TAMRA); Al-X1-X2-N-T-X5-X6-X7-(S-TAMRA); A2-X1-X2-N-T-X5-X6-X7-(5-TAMRA); AS-X1-X2-N-T-X5-X6-X7-(S-TAMRA); A4-X1-X2-N-T-X5-X6-X7-(5-TAMRA); A5-X,- X2-N-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-N-T-X5-X6-X7-(S-TAMRA);
AI-X1-X2-N-P-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-N-P-X5-X6-X7-(S-TAMRA); AS-X1-X2-N-P-X5-X6-X7-(S-TAMRA); A4-X,-X2-N-P-X5-X6-X7-(5-TAMRA); A5-X,- X2-N-P-X5-X6-X7-(5-TAMRA); A6-X, -X2-N-P-X5-X6-X7-(5-TAMRA);
Al-X,-X2-N-Nva-X5-X6-X7-(5-TAMRA); A2-X1-X2-N-Nva-X5-X6-X7-(5- TAMRA); AS-X1-X2-N-NVa-X5-X6-X7-(S-TAMRA); A4-X1-X2-N-Nva-X5-X6-X7-(5- TAMRA); AS-X1-X2-N-NVa-X5-X6-X7-(S-TAMRA)^O-X1-X2-N-NVa-X5-X6-X7-(S- TAMRA);
Al-Xi-X2-N-C(Me)-X5-X6-X7-(S-TAMRA)J AZ-Xi-X2-N-C(Me)-X5-X6-X7-(S- TAMRA); AS-X1-X2-N-C(Me)-X5-X6-X7-(S-TAMRA); A4-Xj -X2-N-(CMe)-X5-X6-X7- (5-TAMRA)J AS-Xi-X2-N-C(Me)-X5-X6-X7-(S-TAMRA); Ao-X1-X2-N-C(Me)-X5-X6- X7-(5-TAMRA);
Al -Xi-X2-N- Y-X5-X6-X7-(5-TAMRA); AZ-X1-X2-N-Y-X5-X6-X7-(S-TAMRA); A3-Xi-X2-N-Y-X5-X6-X7-(5-TAMRA); A4-X,-X2-N-Y-X5-X6-X7-(5-TAMRA); A5-X1- X2-N-Y-X5-X6-X7-(S-TAMRA)J AO-X1-X2-N-Y-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-P-X4-X5-X6-X7-(S-TAMRA); AZ-Xi-X2-P-X4-X5-X6-X7-(S-TAMRA); A3-Xi-X2-P-X4-X5-X6-X7-(5-TAMRA); A4-X,-X2-P-X4-X5-X6-X7-(5-TAMRA); A5-X,- X2-P-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-P-X4-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-P-L-X5-X6-X7-(S-TAMRA); AZ-Xi-X2-P-L-X5-X6-X7-(S-TAMRA); AS-Xi-X2-P-L-X5-X6-X7-(S-TAMRA); A4-Xi-X2-P-L-X5-X6-X7-(5-TAMRA); A5-X,- X2-P-L-X5-X6-X7-(S-TAMRA)J AO-XI-X2-P-L-X5-X6-X7-(S-TAMRA);
ZZ Al-X1-X2-P-F-X5-X6-X7-(S-TAMRA); A2-X,-X2-P-F-X5-X6-X7-(5-TAMRA); AS-Xi-X2-P-F-X5-X6-X7-(S-TAMRA); A4-X,-X2-P-F-X5-X6-X7-(5-TAMRA); A5-X1- X2-P-F-X5-X6-X7-(S-TAMRA)J AO-XI-X2-P-F-X5-X6-X7-(S-TAMRA);
A 1 -Xi-X2-P-Q-X5-X6-X7-(S-TAMRA); A2-X, -X2-P-Q-X5-X6-X7-(S-TAMRA); AS-X1-X2-P-Q-X5-X6-X7-(S-TAMRA); A4-X1-X2-P-Q-X5-X6-X7-(5-TAMRA); A5-X,- X2-P-Q-X5-X6-X7-(S-TAMRA)J AO-X1-X2-P-Q-X5-X6-X7-(S-TAMRA);
Al-X,-X2-P-I-X5-X6-X7-(5-TAMRA); A2-X,-X2-P-I-X5-X6-X7-(5-TAMRA); A3- Xi-X2-P-I-X5-X6-X7-(S-TAMRA); A4-X,-X2-P-I-X5-X6-X7-(5-TAMRA); A5-X1-X2-P-I- X5-X6-X7-(S-TAMRA)J AO-XI-X2-P-I-X5-X6-X7-(S-TAMRA);
Al-X1-X2-P-R-X5-X6-X7-(S-TAMRA); Al-X1-X2-P-R-X5-X6-X7-(S-TAMRA); A3-Xi-X2-P-R-X5-X6-X7-(5-TAMRA); A4-Xi-X2-P-R-X5-X6-X7-(5-TAMRA); A5-X1- X2-P-R-X5-X6-X7-(S-TAMRA)J AO-X1-X2-P-R-X5-X6-X7-(S-TAMRA);
Al-X1-X2-P-G-X5-X6-X7-(S-TAMRA); A2-X!-X2-P-G-X5-X6-X7-(5-TAMRA); AS-X1-X2-P-G-X5-X6-X7-(S-TAMRA); A4-Xi-X2-P-G-X5-X6-X7-(5-TAMRA); A5-X1- X2-P-G-X5-X6-X7-(S-TAMRA); A6-X1 -X2-P-G-X5-X6-X7-(S-TAMRA);
Al -X1-X2-P- A-X5-X6-X7-(5-TAMRA); A2-X!-X2-P-A-X5-X6-X7-(5-TAMRA); A3-Xi-X2-P-A-X5-X6-X7-(5-TAMRA); A4-X1-X2-P-A-X5-X6-X7-(5-TAMRA); A5-X1- X2-P-A-X5-X6-X7-(S-TAMRA); A6-X, -X2-P- A-X5-X6-X7-(5-TAMRA);
AI-X1-X2-P-K-X5-X6-X7-(S-TAMRA)J AI-XI-X2-P-K-X5-X6-X7-(S-TAMRA); A3-Xi-X2-P-K-X5-X6-X7-(5-TAMRA); A4-X1-X2-P-K-X5-X6-X7-(5-TAMRA); A5-X,- X2-P-K-X5-X6-X7-(S-TAMRA)J AO-XI-X2-P-K-X5-X6-X7-(S-TAMRA); Al-X,-X2-P-T-X5-X6-X7-(5-TAMRA); A2-X,-X2-P-T-X5-X6-X7-(5-TAMRA); A3-X,-X2-P-T-X5-X6-X7-(5-TAMRA); A4-X1-X2-P-T-X5-X6-X7-(5-TAMRA); A5-XI- X2-P-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-P-T-X5-X6-X7-(S-TAMRA);
A 1 -X1-X2-P-P-X5-X6-X7-(S-TAMRA); AZ-X1-X2-P-P-X5-X6-X7-(S-TAMRA); AS-Xi-X2-P-P-X5-X6-X7-(S-TAMRA); A4-X,-X2-P-P-X5-X6-X7-(5-TAMRA); A5-Xi- X2-P-P-X5-X6-X7-(S-TAMRA)J AO-XI-X2-P-P-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-P-Nva-X5-X6-X7-(5-TAMRA); A2-X,-X2-P-Nva-X5-X6-X7-(5- TAMRA); AS-Xi-X2-P-NVa-X5-X6-X7-(S-TAMRA); A4-X,-X2-P-Nva-X5-X6-X7-(5- TAMRA); AS-Xi-X2-P-NVa-X5-X6-X7-(S-TAMRA)J Ao-Xi-X2-P-NVa-X5-X6-X7-(S- TAMRA);
Al-X,-X2-P-C(Me)-X5-X6-X7-(5-TAMRA); A2-Xi-X2-P-C(Me)-X5-X6-X7-(5- TAMRA); A3-X, -X2-P-C(Me)-X5-X6-X7-(5-TAMRA); A4-X, -X2-P-(CMe)-X5-X6-X7- (5-TAMRA); AS-Xi-X2-P-C(Me)-X5-X6-X7-(S-TAMRA); AO-X1-X2-P-C(Me)-X5-X6- X7-(5-TAMRA);
Al-X1-X2-P-Y-X5-X6-X7-(S-TAMRA); A2-XrX2-P-Y-X5-X6-X7-(5-TAMRA); A3-X1-X2-P-Y-X5-X6-X7-(5-TAMRA); A4-Xi-X2-P-Y-X5-X6-X7-(5-TAMRA); A5-X1- X2-P-Y-X5-X6-X7-(S-TAMRA)J AO-X1-X2-P-Y-X5-X6-X7-(S-TAMRA);
A1-XI-X2-A-X4-X5-X6-X7-(5-TAMRA); A2-XI-X2-A-X4-X5-X6-X7-(5-TAMRA); A3-Xi-X2-A-X4-X5-X6-X7-(5-TAMRA); A4-Xi-X2-A-X4-X5-X6-X7-(5-TAMRA); A5- XI-X2-A-X4-X5-X6-X7-(S-TAMRA)J AO-XI-X2-A-X4-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-A-L-X5-X6-X7-(S-TAMRA)J Aa-X1-X2-A-L-X5-X6-X7-(S-TAMRA); AS-Xi-X2-A-L-X5-X6-X7-(S-TAMRA); A4-Xi-X2-A-L-X5-X6-X7-(5-TAMRA); A5-X,- X2-A-L-X5-X6-X7-(5-TAMRA); A6-X,-X2-A-L-X5-X6-X7-(5-TAMRA); Al-X,-X2-A-F-X5-X6-X7-(5-TAMRA); A2-X1-X2-A-F-X5-X6-X7-(5-TAMRA); AS-X1-X2-A-F-X5-X6-X7-(S-TAMRA); A4-Xi-X2-A-F-X5-X6-X7-(5-TAMRA); A5-X,- X2-A-F-X5-X6-X7-(S-TAMRA)J AO-XI-X2-A-F-X5-X6-X7-(S-TAMRA);
Al-X1-X2-A-Q-X5-X6-X7-(S-TAMRA); A2-XI-X2-A-Q-X5-X6-X7-(5-TAMRA); A3-Xi-X2-A-Q-X5-X6-X7-(5-TAMRA); A4-X1-X2-A-Q-X5-X6-X7-(5-TAMRA); A5-X1- X2-A-Q-X5-X6-X7-(S-TAMRA); A6-X, -X2-A-Q-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-A-I-X5-X6-X7-(S-TAMRA); A2-X,-X2-A-I-X5-X6-X7-(5-TAMRA); AS-X1-X2-A-I-X5-X6-X7-(S-TAMRA); A4-X1-X2-A-I-X5-X6-X7-(5-TAMRA); A5-Xi- X2-A-I-X5-X6-X7-(5-TAMRA); A6-Xi-X2-A-I-X5-X6-X7-(5-TAMRA);
Al-X1-X2-A-R-X5-X6-X7-(S-TAMRA); A2-X1-X2-A-R-X5-X6-X7-(5-TAMRA); AS-Xi-X2-A-R-X5-X6-X7-(S-TAMRA); A4-X,-X2-A-R-X5-X6-X7-(5-TAMRA); A5-Xr X2-A-R-X5-X6-X7-(5-TAMRA); Ao-X1-X2-A-R-X5-X6-X7-(S-TAMRA);
Al-X1-X2-A-G-X5-X6-X7-(S-TAMRA); A^-X1-X2-A-G-X5-X6-X7-(S-TAMRA); AS-X1-X2-A-G-X5-X6-X7-(S-TAMRA); A4-Xi-X2-A-G-X5-X6-X7-(5-TAMRA); A5-Xr X2-A-G-X5-X6-X7-(S-TAMRA)J AO-XI-X2-A-G-X5-X6-X7-(S-TAMRA);
Al-X1-X2-A-A-X5-X6-X7-(S-TAMRA); A2-XI-X2-A-A-X5-X6-X7-(5-TAMRA); A3-Xi-X2-A-A-X5-X6-X7-(5-TAMRA); A4-X1-X2-A-A-X5-X6-X7-(5-TAMRA); A5-X1- X2-A-A-X5-X6-X7-(S-TAMRA)J AO-XI-X2-A-A-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-A-K-X5-X6-X7-(S-TAMRA); A2-X1-X2-A-K-X5-X6-X7-(5-TAMRA); A3-Xi-X2-A-K-X5-X6-X7-(5-TAMRA); A4-Xi-X2-A-K-X5-X6-X7-(5-TAMRA); A5-X,- X2-A-K-X5-X6-X7-(5-TAMRA); A6-X, -X2- A-K-X5-X6-X7-(5 -TAMRA); Al-X,-X2-A-T-X5-X6-X7-(5-TAMRA); A2-X,-X2-A-T-X5-X6-X7-(5-TAMRA); AS-X1-X2-A-T-X5-X6-X7-(S-TAMRA); A4-X, -X2- A-T-X5-X6-X7-(5 -TAMRA); A5-Xi - X2-A-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-A-T-X5-X6-X7-(S-TAMRA);
AI-XI-X2-A-P-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-A-P-X5-X6-X7-(S-TAMRA); AS-X1-X2-A-P-X5-X6-X7-(S-TAMRA); A4-X,-X2-A-P-X5-X6-X7-(5-TAMRA); A5-Xr X2-A-P-X5-X6-X7-(S-TAMRA)J AO-X1-X2-A-P-X5-X6-X7-(S-TAMRA);
Al-X1-X2-A-NVa-X5-X6-X7-(S-TAMRA); A2-Xi-X2-A-Nva-X5-X6-X7-(5- TAMRA); A3-Xi-X2-A-Nva-X5-X6-X7-(5-TAMRA); A4-X!-X2-A-Nva-X5-X6-X7-(5- TAMRA)J AS-X1-X2-A-NVa-X5-X6-X7-(S-TAMRA)^O-X1-X2-A-NVa-X5-X6-X7-(S- TAMRA);
Al-X1-X2-A-C(Me)-X5-X6-X7-(S-TAMRA); A2-X,-X2-A-C(Me)-X5-X6-X7-(5- TAMRA); A3-Xi-X2-A-C(Me)-X5-X6-X7-(5-TAMRA)j A4-XrX2-A-(CMe)-X5-X6-X7- (5-TAMRA)J AS-Xi-X2-A-C(Me)-X5-X6-X7-(S-TAMRA); AO-X1-X2-A-C(Me)-X5-X6- X7-(5-TAMRA);
Al-Xi-X2-A-Y-X5-X6-X7-(S-TAMRA); A2-XrX2-A- Y-X5-X6-X7-(S-TAMRA); A3-X1-X2-A-Y-X5-X6-X7-(5-TAMRA); A4-Xi-X2-A-Y-X5-X6-X7-(5-TAMRA); A5-Xi- X2-A-Y-X5-X6-X7-(S-TAMRA)J AO-XI-X2-A-Y-X5-X6-X7-(S-TAMRA);
Al -X1-X2-G-X4-X5-X6-X7-(S-TAMRA); A2-Xi -X2-G-X4-X5-X6-X7-(S -T AMRA); AS-X1-X2-G-X4-X5-X6-X7-(S-TAMRA); A4-X,-X2-G-X4-X5-X6-X7-(5-TAMRA); A5- XI-X2-G-X4-X5-X6-X7-(S-TAMRA)^O-XI-X2-G-X4-X5-X6-X7-(S-TAMRA);
AI-XI-X2-G-L-X5-X6-X7-(S-TAMRA)J A^-XI-X2-G-L-X5-X6-X7-(S-TAMRA); A3-XI-X2-G-L-X5-X6-X7-(5-TAMRA); A4-XI-X2-G-L-X5-X6-X7-(5-TAMRA)J A5-X,- X2-G-L-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-L-X5-X6-X7-(S-TAMRA); Al-X1-X2-G-F-X5-X6-X7-(S-TAMRA); A2-XrX2-G-F-X5-X6-X7-(5-TAMRA); AS-X1-X2-G-F-X5-X6-X7-(S-TAMRA); A4-X,-X2-G-F-X5-X6-X7-(5-TAMRA); A5-X,- X2-G-F-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-F-X5-X6-X7-(S-TAMRA);
Al-X1-X2-G-Q-X5-X6-X7-(S-TAMRA); A2-XI-X2-G-Q-X5-X6-X7-(5-TAMRA); AS-X1-X2-G-Q-X5-X6-X7-(S-TAMRA)J A^X1-X2-G-Q-X5-X6-X7-(S-TAMRA)J AS-X1- X2-G-Q-X5-X6-X7-(S-TAMRA)^O-X1-X2-G-Q-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-G-I-X5-X6-X7-(S-TAMRA); A2-X!-X2-G-I-X5-X6-X7-(5-TAMRA); AS-X1-X2-G-I-X5-X6-X7-(S-TAMRA); A4-Xi-X2-G-I-X5-X6-X7-(5-TAMRA); A5-Xr X2-G-I-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-I-X5-X6-X7-(S-TAMRA);
AI-X1-X2-G-R-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-G-R-X5-X6-X7-(S-TAMRA); AS-XI-X2-G-R-X5-X6-X7-(S-TAMRA); A4-X1-X2-G-R-X5-X6-X7-(5-TAMRA); A5-X!- X2-G-R-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-R-X5-X6-X7-(S-TAMRA);
AI-X1-X2-G-G-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-G-G-X5-X6-X7-(S-TAMRA); AS-X1-X2-G-G-X5-X6-X7-(S-TAMRA); A4-X,-X2-G-G-X5-X6-X7-(5-TAMRA); A5-X,- X2-G-G-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-G-X5-X6-X7-(S-TAMRA);
AI-X1-X2-G-A-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-G-A-X5-X6-X7-(S-TAMRA); A3-X1-X2-G-A-X5-X6-X7-(5-TAMRA)j A4-X,-X2-G-A-X5-X6-X7-(5-TAMRA); A5-X1- X2-G-A-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-A-X5-X6-X7-(S-TAMRA)J
Al -Xi-X2-G-K-X5-X6-X7-(S-TAMRA); AZ-X1-X2-G-K-X5-X6-X7-(S-TAMRA); AS-X1-X2-G-K-X5-X6-X7-(S-TAMRA); A4-X,-X2-G-K-X5-X6-X7-(5-TAMRA); A5-Xr X2-G-K-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-K-X5-X6-X7-(S-TAMRA);
Z7 AI-X1-X2-G-T-X5-X6-X7-(S-TAMRA)J AI-X1-X2-G-T-X5-X6-X7-(S-TAMRA); A3-X,-X2-G-T-X5-X6-X7-(5-TAMRA); A4-X,-X2-G-T-X5-X6-X7-(5-TAMRA); A5-XI- X2-G-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-T-X5-X6-X7-(S-TAMRA);
Al-X1-X2-G-P-X5-X6-X7-(S-TAMRA); A2-X1-X2-G-P-X5-X6-X7-(5-TAMRA); AS-X1-X2-G-P-X5-X6-X7-(S-TAMRA); A4-X,-X2-G-P-X5-X6-X7-(5-TAMRA); A5-Xi- X2-G-P-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-P-X5-X6-X7-(S-TAMRA);
Al-X1-X2-G-Nva-X5-X6-X7-(5-TAMRA); A2-X1-X2-G-Nva-X5-X6-X7-(5- TAMRA); AS-X1-X2-G-NVa-X5-X6-X7-(S-TAMRA); A4-X1-X2-G-Nva-X5-X6-X7-(5- TAMRA)J AS-X1-X2-G-NVa-X5-X6-X7-(S-TAMRA)J AO-X1-X2-G-NVa-X5-X6-X7-(S- TAMRA)J
Al-X1-X2-G-C(Me)-X5-X6-X7-(S-TAMRA); A2-X !-X2-G-C(Me)-X5-X6-X7-(S- TAMRA); A3-Xi-X2-G-C(Me)-X5-X6-X7-(5-TAMRA); A4-X1-X2-G-(CMe)-X5-X6-X7- (5-TAMRA)J AS-X1-X2-G-C(Me)-X5-X6-X7-(S-TAMRA); AO-Xi-X2-G-C(Me)-X5-X6- X7-(5-TAMRA);
AI-X1-X2-G-Y-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-G-Y-X5-X6-X7-(S-TAMRA); AS-X1-X2-G-Y-X5-X6-X7-(S-TAMRA); A4-XI-X2-G-Y-X5-X6-X7-(5-TAMRA); A5-X,- X2-G-Y-X5-X6-X7-(S-TAMRA)J AO-XI-X2-G-Y-X5-X6-X7-(S-TAMRA)J
Al-X1-X2-S-X4-X5-X6-X7-(S-TAMRA); A2-Xi-X2-S-X4-X5-X6-X7-(5-TAMRA); AS-X1-X2-S-X4-X5-X6-X7-(S-TAMRA); A4-Xi-X2-S-X4-X5-X6-X7-(5-TAMRA); A5-X,- X2-S-X4-X5-X6-X7-(S-TAMRA)^O-X1-X2-S-X4-X5-X6-X7-(S-TAMRA);
Al-X1-X2-S-L-X5-X6-X7-(S-TAMRA)J Aa-Xi-X2-S-L-X5-X6-X7-(S-TAMRA); AS-X1-X2-S-L-X5-X6-X7-(S-TAMRA); A4-X1-X2-S-L-X5-X6-X7-(5-TAMRA); A5-Xi- X2-S-L-X5-X6-X7-(S-TAMRA)J AO-X1-X2-S-L-X5-X6-X7-(S-TAMRA); Al-X1-X2-S-F-X5-X6-X7-(S-TAMRA); A2-X1-X2-S-F-X5-X6-X7-(5-TAMRA); A3-X,-X2-S-F-X5-X6-X7-(5-TAMRA); A4-X,-X2-S-F-X5-X6-X7-(5-TAMRA); A5-X,- X2-S-F-X5-X6-X7-(S-TAMRA)J AO-X1-X2-S-F-X5-X6-X7-(S-TAMRA);
AI-X1-X2-S-Q-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-S-Q-X5-X6-X7-(S-TAMRA); AS-X1-X2-S-Q-X5-X6-X7-(S-TAMRA); A4-X,-X2-S-Q-X5-X6-X7-(5-TAMRA); A5-X,- X2-S-Q-X5-X6-X7-(S-TAMRA)J AO-XI-X2-S-Q-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-S-I-X5-X6-X7-(5-TAMRA); A2-X1-X2-S-I-X5-X6-X7-(5-TAMRA); A3- X1-X2-S-I-X5-X6-X7-(S-TAMRA); A4-Xi-X2-S-I-X5-X6-X7-(5-TAMRA); AS-X1-X2-S-I- X5-X6-X7-(S-TAMRA)J AO-XI-X2-S-I-X5-X6-X7-(S-TAMRA);
Al-X1-X2-S-R-X5-X6-X7-(S-TAMRA); A2-X,-X2-S-R-X5-X6-X7-(5-TAMRA); AS-Xi-X2-S-R-X5-X6-X7-(S-TAMRA); A4-Xi-X2-S-R-X5-X6-X7-(5-TAMRA); A5-X1- X2-S-R-X5-X6-X7-(5-TAMRA); A6-X, -X2-S-R-X5-X6-X7-(5-TAMRA);
Al-X1-X2-S-G-X5-X6-X7-(S-TAMRA); A2-Xi-X2-S-G-X5-X6-X7-(5-TAMRA); A3-X,-X2-S-G-X5-X6-X7-(5-TAMRA); A4-X,-X2-S-G-X5-X6-X7-(5-TAMRA); A5-Xi- X2-S-G-X5-X6-X7-(S-TAMRA)J AO-X1-X2-S-G-X5-X6-X7-(S-TAMRA);
Al-X1-X2-S-A-X5-X6-X7-(S-TAMRA); A2-XI-X2-S-A-X5-X6-X7-(5-TAMRA); AS-X1-X2-S-A-X5-X6-X7-(S-TAMRA); A4-X,-X2-S-A-X5-X6-X7-(5-TAMRA); A5-X,- X2-S-A-X5-X6-X7-(S-TAMRA)J AO-X1-X2-S-A-X5-X6-X7-(S-TAMRA);
AI-XI-X2-S-K-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-S-K-X5-X6-X7-(S-TAMRA); AS-XI-X2-S-K-X5-X6-X7-(S-TAMRA); A4-X,-X2-S-K-X5-X6-X7-(5-TAMRA); A5-Xr X2-S-K-X5-X6-X7-(S-TAMRA)J AO-XI-X2-S-K-X5-X6-X7-(S-TAMRA)J Al-X1-X2-S-T-X5-X6-X7-(S-TAMRA); A2-X,-X2-S-T-X5-X6-X7-(5-TAMRA); AS-X1-X2-S-T-X5-X6-X7-(S-TAMRA); A4-X1-X2-S-T-X5-X6-X7-(5-TAMRA); A5-X,- X2-S-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-S-T-X5-X6-X7-(S-TAMRA);
AI-X1-X2-S-P-X5-X6-X7-(S-TAMRA)J AZ-XI-X2-S-P-X5-X6-X7-(S-TAMRA); AS-X1-X2-S-P-X5-X6-X7-(S-TAMRA); A4-Xi-X2-S-P-X5-X6-X7-(5-TAMRA); A5-Xr X2-S-P-X5-X6-X7-(S-TAMRA^ AO-X1-X2-S-P-X5-X6-X7-(S-TAMRA);
Al -X1-X2-S-NVa-X5-X6-X7-(S-TAMRA); A2-X!-X2-S-Nva-X5-X6-X7-(5- TAMRA); AS-X1-X2-S-NVa-X5-X6-X7-(S-TAMRA); A4-X1-X2-S-Nva-X5-X6-X7-(5- TAMRA); AS-X1-X2-S-NVa-X5-X6-X7-(S-TAMRA); A6-X,-X2-S-Nva-X5-X6-X7-(5- TAMRA);
Al-X1-X2-S-C(Me)-X5-X6-X7-(S-TAMRA); A2-X,-X2-S-C(Me)-X5-X6-X7-(5- TAMRA); AS-X1-X2-S-C(Me)-X5-X6-X7-(S-TAMRA); A4-Xi -X2-S-(CMe)-X5-X6-X7- (5-TAMRA)J AS-Xi-X2-S-C(Me)-X5-X6-X7-(S-TAMRA); AO-X1-X2-S-C(Me)-X5-X6- X7-(5-TAMRA);
AI-XI-X2-S-Y-X5-X6-X7-(S-TAMRA)J AI-X1-X2-S-Y-X5-X6-X7-(S-TAMRA); A3-Xi -X2-S- Y-X5-X6-X7-(S-TAMRA); A4-XI -X2-S- Y-X5-X6-X7-(S-TAMRA); A5-XI - X2-S-Y-X5-X6-X7-(S-TAMRA)J AO-XI-X2-S-Y-X5-X6-X7-(S-TAMRA);
AI-X1-X2-L-X4-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-L-X4-X5-X6-X7-(S-TAMRA); AS-XI-X2-L-X4-X5-X6-X7-(S-TAMRA); A4-X,-X2-L-X4-X5-X6-X7-(5-TAMRA)J A5-X,- X2-L-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-L-X4-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-L-L-X5-X6-X7-(S-TAMRA)J A2-X,-X2-L-L-X5-X6-X7-(5-TAMRA)J A3-X,-X2-L-L-X5-X6-X7-(5-TAMRA); A4-X,-X2-L-L-X5-X6-X7-(5-TAMRA)j A5-X1- X2-L-L-X5-X6-X7-(5-TAMRA); A6-Xi-X2-L-L-X5-X6-X7-(5-TAMRA); Al-X1-X2-L-F-X5-X6-X7-(S-TAMRA); A2-Xi-X2-L-F-X5-X6-X7-(5-TAMRA); A3-X,-X2-L-F-X5-X6-X7-(5-TAMRA); A4-X,-X2-L-F-X5-X6-X7-(5-TAMRA); A5-X,- X2-L-F-X5-X6-X7-(S-TAMRA)S AO-XI-X2-L-F-X5-X6-X7-(S-TAMRA);
AI-X1-X2-L-Q-X5-X6-X7-(S-TAMRA)J AI-X1-X2-L-Q-X5-X6-X7-(S-TAMRA); AS-X1-X2-L-Q-X5-X6-X7-(S-TAMRA); A4-X,-X2-L-Q-X5-X6-X7-(5-TAMRA); A5-X1- X2-L-Q-X5-X6-X7-(5-TAMRA); A6-X, -X2-L-Q-X5-X6-X7-(5-TAMRA);
AI-XI-X2-L-I-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-L-I-X5-X6-X7-(S-TAMRA); AS-XI-X2-L-I-X5-X6-X7-(S-TAMRA); A4-Xi-X2-L-I-X5-X6-X7-(5-TAMRA); A5-XrX2- L-I-X5-X6-X7-(S-TAMRA)J AO-X1-X2-L-I-X5-X6-X7-(S-TAMRA);
Al-X1-X2-L-R-X5-X6-X7-(S-TAMRA); A2-XI-X2-L-R-X5-X6-X7-(5-TAMRA); AS-X1-X2-L-R-X5-X6-X7-(S-TAMRA); A4-Xi-X2-L-R-X5-X6-X7-(5-TAMRA); A5-Xi- X2-L-R-X5-X6-X7-(S-TAMRA)J AO-XI-X2-L-R-X5-X6-X7-(S-TAMRA);
A1-XI-X2-L-G-X5-X6-X7-(5-TAMRA); A2-XI-X2-L-G-X5-X6-X7-(5-TAMRA); AS-X1-X2-L-G-X5-X6-X7-(S-TAMRA); A4-X,-X2-L-G-X5-X6-X7-(5-TAMRA); A5-X,- X2-L-G-X5-X6-X7-(5-TAMRA) j A6-X, -X2-L-G-X5-X6-X7-(5-TAMRA)j
Al-Xi-X2-L-A-X5-X6-X7-(5-TAMRA); A2-X1-X2-L-A-X5-X6-X7-(5-TAMRA)j A3-XI-X2-L-A-X5-X6-X7-(5-TAMRA); A4-XI-X2-L-A-X5-X6-X7-(5-TAMRA); A5-X1- X2-L- A-X5-X6-X7-(5-TAMRA); A6-X. -X2-L- A-X5-X6-X7-(5-TAMRA);
A 1 -X1-X2-L-K-X5-X6-X7-(S-TAMRA)J A2-X,-X2-L-K-X5-X6-X7-(5-TAMRA); A3-Xi-X2-L-K-X5-X6-X7-(5-TAMRA)j A4-X,-X2-L-K-X5-X6-X7-(5-TAMRA); A5-Xi- X2-L-K-X5-X6-X7-(S-TAMRA)J AO-X1-X2-L-K-X5-X6-X7-(S-TAMRA); Al-Xi-X2-L-T-X5-X6-X7-(5-TAMRA); A2-X,-X2-L-T-X5-X6-X7-(5-TAMRA); A3-X,-X2-L-T-X5-X6-X7-(5-TAMRA); A4-Xi-X2-L-T-X5-X6-X7-(5-TAMRA); A5-X,- X2-L-T-X5-X6-X7-(S-TAMRA)J AO-XI-X2-L-T-X5-X6-X7-(S-TAMRA);
AI-XI-X2-L-P-X5-X6-X7-(S-TAMRA)5 AI-X1-X2-L-P-X5-X6-X7-(S-TAMRA); AS-X1-X2-L-P-X5-X6-X7-(S-TAMRA); A4-X,-X2-L-P-X5-X6-X7-(5-TAMRA); A5-X,- X2-L-P-X5-X6-X7-(S-TAMRA)J AO-X1-X2-L-P-X5-X6-X7-(S-TAMRA);
A 1 -X, -X2-L-Nva-X5-X6-X7-(5-TAMRA); A2-X, -X2-L-Nva-X5-X6-X7-(5- TAMRA); AB-X1-X2-L-NVa-X5-X6-X7-(S-TAMRA); A4-X1-X2-L-Nva-X5-X6-X7-(5- TAMRA)J AS-Xi-X2-L-NVa-X5-X6-X7-(S-TAMRA)J AO-X1-X2-L-NVa-X5-X6-X7-(S- TAMRA);
Al-Xi-X2-L-C(Me)-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-L-C(Me)-X5-X6-X7-(S- TAMRA)J AS-X1-X2-L-C(Me)-X5-X6-X7-(S-TAMRA); A4-XrX2-L-(CMe)-X5-X6-X7- (5-TAMRA); AS-Xi-X2-L-C(Me)-X5-X6-X7-(S-TAMRA); AO-X1-X2-L-C(Me)-X5-X6- X7-(5-TAMRA)j
AI-X1-X2-L-Y-X5-X6-X7-(S-TAMRA)J AI-X1-X2-L-Y-X5-X6-X7-(S-TAMRA)J AS-X1-X2-L-Y-X5-X6-X7-(S-TAMRA); A4-X,-X2-L-Y-X5-X6-X7-(5-TAMRA); A5-XI- X2-L-Y-X5-X6-X7-(S-TAMRA); A6-X1 -X2-L-Y-X5-X6-X7-(S-TAMRA);
Al-X1-X2-V-X4-X5-X6-X7-(S-TAMRA); A2-X,-X2- V-X4-X5-X6-X7-(S-T AMRA); AS-X1-X2-V-X4-X5-X6-X7-(S-TAMRA); A4-X,-X2- V-X4-X5-X6-X7-(S-TAMRA); A5- X1-X2-V-X4-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-X4-X5-X6-X7-(S-TAMRA);
AI-X1-X2-V-L-X5-X6-X7-(S-TAMRA)J AI-X1-X2-V-L-X5-X6-X7-(S-TAMRA); AS-X1-X2-V-L-X5-X6-X7-(S-TAMRA); A4-XrX2- V-L-X5-X6-X7-(S-TAMRA)J A5-XI- X2-V-L-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-L-X5-X6-X7-(S-TAMRA)J Al-X1-X2-V-F-X5-X6-X7-(S-TAMRA); A2-X,-X2-V-F-X5-X6-X7-(5-TAMRA); A3-Xi-X2-V-F-X5-X6-X7-(5-TAMRA); A4-X,-X2-V-F-X5-X6-X7-(5-TAMRA); A5-X,- X2- V-F-X5-X6-X7-(5-TAMRA); AO-X1-X2-V-F-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-V-Q-X5-X6-X7-(S-TAMRA); A2-X,-X2- V-Q-X5-X6-X7-(S-TAMRA); A3-X,-X2-V-Q-X5-X6-X7-(5-TAMRA); A4-Xi-X2-V-Q-X5-X6-X7-(5-TAMRA); A5-X,- X2-V-Q-X5-X6-X7-(S-TAMRA)5 AO-X1-X2-V-Q-X5-X6-X7-(S-TAMRA);
Al-X,-X2-V-I-X5-X6-X7-(5-TAMRA); A2-X,-X2-V-I-X5-X6-X7-(5-TAMRA); AS-X1-X2-V-I-X5-X6-X7-(S-TAMRA); A4-X1-X2-V-I-X5-X6-X7-(5-TAMRA); A5-Xr X2-V-I-X5-X6-X7-(5-TAMRA); A6-Xi-X2-V-I-X5-X6-X7-(5-TAMRA);
Al-X1-X2-V-R-X5-X6-X7-(S-TAMRA); A2-X,-X2-V-R-X5-X6-X7-(5-TAMRA); A3-Xi -X2-V-R-X5-X6-X7-(S-TAMRA); A4-X, -X2- V-R-X5-X6-X7-(5-TAMRA); A5-Xi - X2-V-R-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-R-X5-X6-X7-(S-TAMRA);
Al-Xi-X2- V-G-X5-X6-X7-(5-TAMRA); A2-X,-X2-V-G-X5-X6-X7-(5-TAMRA); AS-X1-X2-V-G-X5-X6-X7-(S-TAMRA); A4-X,-X2-V-G-X5-X6-X7-(5-TAMRA); A5-X,- X2-V-G-X5-X6-X7-(S-TAMRA)J AO-XI-X2-V-G-X5-X6-X7-(S-TAMRA);
AI-XI-X2-V-A-X5-X6-X7-(S-TAMRA)J A^-X1-X2-V-A-X5-X6-X7-(S-TAMRA); A3-XI-X2-V-A-X5-X6-X7-(5-TAMRA); A4-X,-X2-V-A-X5-X6-X7-(5-TAMRA); A5-XI- X2-V-A-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-A-X5-X6-X7-(S-TAMRA);
A 1 -X1-X2-V-K-X5-X6-X7-(S-TAMRA); A2-X] -X2-V-K-X5-X6-X7-(S-TAMRA); A3-Xi-X2-V-K-X5-X6-X7-(5-TAMRA); A4-X,-X2-V-K-X5-X6-X7-(5-TAMRA); A5-Xi- X2-V-K-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-K-X5-X6-X7-(S-TAMRA); Al-X1-X2-V-T-X5-X6-X7-(S-TAMRA); A2-X1-X2-V-T-X5-X6-X7-(5-TAMRA); A3-X1-X2-V-T-X5-X6-X7-(5-TAMRA); A4-Xi-X2-V-T-X5-X6-X7-(5-TAMRA); A5-X1- X2-V-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-T-X5-X6-X7-(S-TAMRA);
Al -X1-X2- V-P-X5-X6-X7-(5-TAMRA); Al-X1-X2-V-P-X5-X6-X7-(S-TAMRA); AS-X1-X2-V-P-X5-X6-X7-(S-TAMRA); A4-X!-X2-V-P-X5-X6-X7-(5-TAMRA); A5-X,- X2-V-P-X5-X6-X7-(S-TAMRA)J AO-XI-X2-V-P-X5-X6-X7-(S-TAMRA);
Al-X1-X2-V-NVa-X5-X6-X7-(S-TAMRA)J AZ-Xi-X2-V-NVa-X5-X6-X7-(S- TAMRA); A3-Xj-X2-V-Nva-X5-X6-X7-(5-TAMRA); A4-XrX2- V-Nva-X5-X6-X7-(5- TAMRA)J AS-X1-X2-V-NVa-X5-X6-X7-(S-TAMRA)J Ao-Xi-X2-V-NVa-X5-X6-X7-(S- TAMRA);
Al-X1-X2-V-C(Me)-X5-X6-X7-(S-TAMRA); Al-X1-X2-V-C(Me)-X5-X6-X7-(S- TAMRA); A3-Xi-X2-V-C(Me)-X5-X6-X7-(5-TAMRA); A4-XrX2- V-(CMe)-X5-X6-X7- (5-TAMRA)J AS-Xi-X2-V-C(Me)-X5-X6-X7-(S-TAMRA); Ao-X1-X2- V-C(Me)-X5-X6- X7-(5-TAMRA);
AI-XI-X2-V-Y-X5-X6-X7-(S-TAMRA)J AI-XI-X2-V-Y-X5-X6-X7-(S-TAMRA)J A3-Xi-X2-V-Y-X5-X6-X7-(5-TAMRA)j A4-X1-X2-V-Y-X5-X6-X7-(5-TAMRA)j A5-X1- X2-V-Y-X5-X6-X7-(S-TAMRA)J AO-X1-X2-V-Y-X5-X6-X7-(S-TAMRA)J
Al -X1-X2-K-X4-X5-X6-X7-(S-TAMRA)J A2-Xi-X2-K-X4-X5-X6-X7-(5-T AMRA); AS-Xi-X2-K-X4-X5-X6-X7-(S-TAMRA); A4-X1-X2-K-X4-X5-X6-X7-(5-TAMRA); A5- XI-X2-K-X4-X5-X6-X7-(S-TAMRA)J AO-XI-X2-K-X4-X5-X6-X7-(S-TAMRA);
AI-XI-X2-K-L-X5-X6-X7-(S-TAMRA)J AI-X1-X2-K-L-X5-X6-X7-(S-TAMRA); AS-XI-X2-K-L-X5-X6-X7-(S-TAMRA); A4-XI-X2-K-L-X5-X6-X7-(5-TAMRA); A5-X,- X2-K-L-X5-X6-X7-(S-TAMRA); A6-X1 -X2-K-L-X5-X6-X7-(5-TAMRA)j A 1 -Xi -X2-K-F-X5-X6-X7-(S-TAMRA); A2-X, -X2-K-F-X5-X6-X7-(S-TAMRA); AS-Xi-X2-K-F-X5-X6-X7-(S-TAMRA); A4-X,-X2-K-F-X5-X6-X7-(5-TAMRA); A5-Xt- X2-K-F-X5-X6-X7-(S-TAMRA)J AO-X1-X2-K-F-X5-X6-X7-(S-TAMRA);
AI-X1-X2-K-Q-X5-X6-X7-(S-TAMRA)J AI-X1-X2-K-Q-X5-X6-X7-(S-TAMRA); AS-X1-X2-K-Q-X5-X6-X7-(S-TAMRA); A4-XI-X2-K-Q-X5-X6-X7-(5-TAMRA); A5-XI- X2-K-Q-X5-X6-X7-(S-TAMRA)J AO-X1-X2-K-Q-X5-X6-X7-(S-TAMRA);
AI-X1-X2-K-I-X5-X6-X7-(S-TAMRA)J AI-XI-X2-K-I-X5-X6-X7-(S-TAMRA); A3-XI-X2-K-I-X5-X6-X7-(5-TAMRA); A4-X,-X2-K-I-X5-X6-X7-(5-TAMRA); A5-XI- X2-K-I-X5-X6-X7-(S-TAMRA); AO-Xi-X2-K-I-X5-X6-X7-(S-TAMRA);
Al-X1-X2-K-R-X5-X6-X7-(S-TAMRA); A^-X1-X2-K-R-X5-X6-X7-(S-TAMRA); AS-X1-X2-K-R-X5-X6-X7-(S-TAMRA); A4-X1-X2-K-R-X5-X6-X7-(5-TAMRA); A5-Xi- X2-K-R-X5-X6-X7-(S-TAMRA); AO-X1-X2-K-R-X5-X6-X7-(S-TAMRA);
AI-X1-X2-K-G-X5-X6-X7-(S-TAMRA)J AZ-X1-X2-K-G-X5-X6-X7-(S-TAMRA); A3-X1-X2-K-G-X5-X6-X7-(5-TAMRA)J A4-XI-X2-K-G-X5-X6-X7-(5-TAMRA); A5-XI- X2-K-G-X5-X6-X7-(S-TAMRA)J AO-XI-X2-K-G-X5-X6-X7-(S-TAMRA)J
Al-X1-X2-K-A-X5-X6-X7-(S-TAMRA); A2-XI-X2-K-A-X5-X6-X7-(5-TAMRA); AS-Xi-X2-K-A-X5-X6-X7-(S-TAMRA); A4-Xi-X2-K-A-X5-X6-X7-(5-TAMRA); A5-Xi- X2-K-A-X5-X6-X7-(S-TAMRA)J AO-X1-X2-K-A-X5-X6-X7-(S-TAMRA);
AI-X1-X2-K-K-X5-X6-X7-(S-TAMRA)J AI-X1-X2-K-K-X5-X6-X7-(S-TAMRA); AS-XI-X2-K-K-X5-X6-X7-(S-TAMRA); A4-X,-X2-K-K-X5-X6-X7-(5-TAMRA); A5-X,- X2-K-K-X5-X6-X7-(S-TAMRA); A6-X, -X2-K-K-X5-X6-X7-(S-TAMRA); A 1 -X1-X2-K-T-X5-X6-X7-(S-TAMRA); A2-X, -X2-K-T-X5-X6-X7-(S-TAMRA); A3-X,-X2-K-T-X5-X6-X7-(5-TAMRA); A4-XI-X2-K-T-X5-X6-X7-(5-TAMRA); A5-X1- X2-K-T-X5-X6-X7-(S-TAMRA)J AO-X1-X2-K-T-X5-X6-X7-(S-TAMRA);
AI-X1-X2-K-P-X5-X6-X7-(S-TAMRA)J AI-X1-X2-K-P-X5-X6-X7-(S-TAMRA); A3-XI -X2-K-P-X5-X6-X7-(S-TAMRA); A4-X, -X2-K-P-X5-X6-X7-(5-TAMRA); A5-XI - X2-K-P-X5-X6-X7-(S-TAMRA)J AO-X1-X2-K-P-X5-X6-X7-(S-TAMRA);
Al-Xi-X2-K-NVa-X5-X6-X7-(S-TAMRA)J Al-X1-X2-K-NVa-X5-X6-X7-(S- TAMRA); AS-Xi-X2-K-NVa-X5-X6-X7-(S-TAMRA); A4-X,-X2-K-Nva-X5-X6-X7-(5- TAMRA); AS-Xi-X2-K-NVa-X5-X6-X7-(S-TAMRA); Ao-Xj-X2-K-NVa-X5-X6-X7-(S- TAMRA);
Al-X1-X2-K-C(Me)-X5-X6-X7-(S-TAMRA); A2-Xi-X2-K-C(Me)-X5-X6-X7-(5- TAMRA); AS-X1-X2-K-C(Me)-X5-X6-X7-(S-TAMRA)J A4-X!-X2-K-(CMe)-X5-X6-X7- (5-TAMRA)J AS-Xi-X2-K-C(Me)-X5-X6-X7-(S-TAMRA); AO-Xi-X2-K-C(Me)-X5-X6- X7-(5-TAMRA);
AI-X1-X2-K-Y-X5-X6-X7-(S-TAMRA)J AI-XI-X2-K-Y-X5-X6-X7-(S-TAMRA); AS-XI-X2-K-Y-X5-X6-X7-(S-TAMRA); A4-X1-X2-K-Y-X5-X6-X7-(5-TAMRA); A5-Xr X2-K- Y-X5-X6-X7-(5-TAMRA); AO-Xi-X2-K-Y-X5-X6-X7-(S-TAMRA);

Claims

CLAIMS:
1. A protease substrate selected from a group of protease substrates consisting of:
C 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(B 1 )-OH; C 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(B2)-OH; C 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; Cl-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ue-Val-Gln-Lys(B4)-OH; Cl-γ-Abu-Ser- GIn-ASn-Ty^PrO-IIe-VaI-GIn-LyS(BS)-OH5 Cl-Y-AbU-SCr-GIn-ASn-TyT-PrO-IIe-VaI- Gln-Lys(B6)-OH;
C2-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(B 1 )-OH; C2-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B2)-OH; C2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C2-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; C2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(B6)-OH;
C3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(B I)-OH; C3-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B2)-OH; C3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C3-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; C3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(B6)-OH;
C4-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(B 1 )-OH; C4-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(B2)-OH; C4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C4-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; C4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-πe-Val- Gln-Lys(B6)-OH; CS-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(B I)-OH; C5-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ue-Val-Gln-Lys(B2)-OH; C5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(B3)-OH; C5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C5-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B5)-OH; C5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(B6)-OH;
C6-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(B 1 )-OH; C6-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile- VaI-GIn-LyS(Ba)-OHj Co-Y-AbU-SeI-GIn-ASn-TyT-PrO-IIe-VaI-GIn- Lys(B3)-OH; C6-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(B4)-OH; C6-γ-Abu-Ser- GIn-ASn-TyT-PrO-IIe-VaI-GIn-LyS(BS)-OHj Co-Y-AbU-SCr-GIn-ASn-TyT-PrO-IIe-VaI- Gln-Lys(B6)-OH.
2. A protease substrate selected from a group of protease substrates consisting of:
B 1 -γ- Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B 1 -γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln- Lys(C3)-0H; B 1 -γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(C4)-OH; B 1 -γ-Abu-Ser- GIn- Asn-Tyr-Pro-Ile- Val-Gln-Lys(C5)-0H; B 1 -γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val- Gln-Lys(C6)-OH;
B2-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B2-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH; B2-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C5)-OH; B2-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(C6)-0H; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B3-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH; B3-γ-Abu-Ser- Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C5)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val- Gln-Lys(C6)-OH;
B4-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(C I)-OH; B3-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B3-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH; B3-γ-Abu-Ser- GIn-ASn-TyT-PrO-IIe-VaI-GIn-LyS(CS)-OHj BS-Y-AbU-SeF-GIn-ASn-TyT-PrO-IIe-VaI- Gln-Lys(C6)-OH;
BS-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile- Val-Gln-Lys(C 1 )-OH; B5-γ-Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B5-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH; B5-γ-Abu-Seτ- GIn-ASn-TyT-PrO-IIe-VaI-GIn-LyS(CS)-OHi BS-Y-AbU-SeT-GIn-ASn-TyT-PrO-IIe-VaI- Gln-Lys(C6)-OH;
B6-γ-Abu-Ser-Gln-Asn-Tyτ-Pro-Ile-Val-Gln-Lys(C 1 )-0H; B6-γ- Abu-Ser-Gln- Asn-Tyr-Pro-Ile-Val-Gln-Lys(C2)-OH; B6-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln- Lys(C3)-OH; B6-γ-Abu-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(C4)-OH; B6-γ-Abu-Ser- GIn-ASn-TyT-PrO-IIe-VaI-GIn-LyS(CS)-OHj Bo-Y-AbU-SeT-GIn-ASn-TyT-PrO-IIe-VaI- Gln-Lys(C6)-0H.
3. A protease substrate selected from a group of protease substrates consisting of:
Al -KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)-AK-NH2; A2-KGP-Cha-Abu-Smc- HAr-K(5-TAMRA)-AK-NH2; A3-KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)-AK-NH2; A4-KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)-AK-NH2; A5-KGP-Cha-Abu-Smc-HAr- K(5-TAMRA)-AK-NH2; A6-KGP-Cha-Abu-Smc-HAr-K(5-TAMRA)-AK-NH2.
PCT/US2010/001268 2009-05-01 2010-04-30 Optimal fret pairs and related methods WO2010126600A1 (en)

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