A general method for the preparation of internally quenched fluorogenic protease substrates using solid-phase peptide synthesis

J Med Chem. 1992 Oct 16;35(21):3727-30. doi: 10.1021/jm00099a001.

Abstract

A general scheme for obtaining a fluorescent donor/acceptor peptide substrate via solid-phase synthesis methodology is presented. The key feature of this method is the design of a glutamic acid derivative that has been modified on the carboxyl side chain with a 5-[(2'-aminoethyl)-amino]naphthelenesulfonic acid (EDANS) to create a fluorescent donor moiety that can be incorporated near the C-terminus of the peptide substrate. The corresponding fluorescent acceptor group containing a 4-[[4-(dimethylamino)phenyl]azo]benzoic acid (DABCYL) can then be attached to the resin-bound peptide at the N-terminus while all side-chain groups are still fully protected. Substrates for renin and HIV proteinase are synthesized as examples.

MeSH terms

  • Amino Acid Sequence
  • Endopeptidases / metabolism*
  • Fluorescent Dyes / chemical synthesis*
  • HIV Protease / metabolism
  • HIV-1 / enzymology
  • Molecular Sequence Data
  • Naphthalenesulfonates / chemical synthesis
  • Naphthalenesulfonates / metabolism
  • Peptides / chemical synthesis*
  • Renin / metabolism
  • Substrate Specificity
  • p-Dimethylaminoazobenzene / analogs & derivatives

Substances

  • Fluorescent Dyes
  • Naphthalenesulfonates
  • Peptides
  • 5-((2-aminoethyl)amino)naphthalene-1-sulfonic acid
  • 4-(4-dimethylaminophenylazo)benzoic acid
  • p-Dimethylaminoazobenzene
  • Endopeptidases
  • HIV Protease
  • Renin