A highly efficient method for site-specific modification of unprotected peptides after chemical synthesis

Steven J. Bark, Sandra Schmid, Klaus M. Hahn

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

We have developed a highly efficient method for the site-specific attachment of biophysical probes to unprotected peptides after chemical synthesis. This methodology takes advantage of the selective reactivity of an N-methylaminooxy amino acid that is appropriately protected for direct incorporation during solid-phase peptide synthesis. The functional N- methylaminooxy group is unmasked using normal peptide cleavage conditions and is capable of selective reaction with activated N-hydroxysuccinimide esters in the presence of cysteine, lysine and the amino terminus, as demonstrated in model peptides and test proteins. Selective labeling can be accomplished after synthesis using commercially available or chemically sensitive probes. This technology is compatible with the synthesis of C(α)-thioester- containing peptides and amide-forming ligations, required steps for the synthesis of proteins by either total chemical synthesis or expressed protein ligation. The N-methylaminooxy amino acid can be introduced into different sites by parallel peptide synthesis to generate a polypeptide analogue family with each member possessing a single specifically labeled site. This enables the synthesis of optimized biosensors through combinatorial screening of different attachment sites for maximal response and minimal perturbation of biological activity.

Original languageEnglish (US)
Pages (from-to)3567-3573
Number of pages7
JournalJournal of the American Chemical Society
Volume122
Issue number15
DOIs
StatePublished - Apr 19 2000

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Peptides
Ligation
Proteins
Amino acids
Amino Acids
Solid-Phase Synthesis Techniques
Biosensing Techniques
Amides
Lysine
Cysteine
Polypeptides
Esters
Bioactivity
Biosensors
Labeling
Technology
Screening

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

A highly efficient method for site-specific modification of unprotected peptides after chemical synthesis. / Bark, Steven J.; Schmid, Sandra; Hahn, Klaus M.

In: Journal of the American Chemical Society, Vol. 122, No. 15, 19.04.2000, p. 3567-3573.

Research output: Contribution to journalArticle

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