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A synthetic GFP-like chromophore undergoes base-catalyzed autoxidation into acylimine red form

J Org Chem. 2011 Apr 15;76(8):2782-91. doi: 10.1021/jo200150b. Epub 2011 Mar 22.

Abstract

Fluorescent proteins are widely used in modern experimental biology, but much controversy exists regarding details of maturation of different types of their chromophores. Here we studied possible mechanisms of DsRed-type red chromophore formation using synthetic biomimetic GFP-like chromophores, bearing an acylamino substituent, corresponding to an amino acid residue at position 65. We have shown these model compounds to readily react with molecular oxygen to produce a highly unstable DsRed-like acylimine, isolated in the form of stable derivatives. Under the same aerobic conditions an unusual red-shifted imide chromophore--a product of 4-electron oxidation of Gly65 residue--is formed. Our data showed that GFP chromophore is prone to autoxidation at position 65 Cα by its chemical nature with basic conditions being the only key factor required.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkalies / chemistry
  • Amino Acid Substitution
  • Catalysis
  • Cyclization
  • Green Fluorescent Proteins / chemical synthesis*
  • Green Fluorescent Proteins / metabolism
  • Imines / chemistry
  • Luminescent Proteins / chemistry*
  • Molecular Mimicry
  • Molecular Probes / chemical synthesis*
  • Oxidation-Reduction
  • Oxygen / metabolism

Substances

  • Alkalies
  • Imines
  • Luminescent Proteins
  • Molecular Probes
  • fluorescent protein 583
  • Green Fluorescent Proteins
  • Oxygen