Peptide biotinylation with amine-reactive esters: Differential side chain reactivity

Brian T. Miller, Thomas J. Collins, Mark E. Rogers, Alexander Kurosky

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

N-hydroxysuccinimide (NHS) esters of biotin are reported to react specifically with amino groups of peptides and proteins. However, we have found that these reagents can readily acylate other functional groups in specific peptide sequences under relatively mild conditions. We have extended our inquiry of sequence-dependent acylation by evaluating the reactivity of a variety of commonly employed biotinylation reagents typically used for amino group modification. These included the p-nitrophenyl ester of biotin, NHS- esters of biotin containing aminohexanoic acid spacer arms, and a sulfonated NHS-biotin ester that contained a disulfide bond within its spacer. The decapeptide [D-Lys6]gonadotropin releasing hormone was employed as a model peptide. Reaction products were characterized by high-performance liquid chromatography, amino acid compositional analysis, reaction with hydroxylamine, and mass spectrometry. In addition to the O-acylation of Ser4 and Tyr5 in this peptide, we have also identified a novel biotinylation of the Arg8 side chain.

Original languageEnglish (US)
Pages (from-to)1585-1595
Number of pages11
JournalPeptides
Volume18
Issue number10
DOIs
StatePublished - Dec 1 1997

Keywords

  • Aminohexanoic acid
  • Arginine modification
  • Biotin-p-nitrophenyl ester
  • Biotinylation [D-Lys]GnRH
  • NHS-biotin esters
  • O-acylation
  • Solvent effects

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Endocrinology
  • Cellular and Molecular Neuroscience

Fingerprint Dive into the research topics of 'Peptide biotinylation with amine-reactive esters: Differential side chain reactivity'. Together they form a unique fingerprint.

Cite this