Boc-Cys(Npys)-OH (BCNP): an appropriate reagent for the identification of T cell epitopes in cystine and/or cysteine-containing proteins

J Immunol Methods. 1994 May 2;171(1):65-71. doi: 10.1016/0022-1759(94)90229-1.

Abstract

Some T cell epitopes become inactive when their thiols are blocked with various irreversible reagents (Régnier-Vigouroux, 1988; Maillère, 1992; Maillère et al., 1993). Blocking protein and peptide thiols with BCNP (Boc-Cys(Npys)-OH) constitutes a most appropriate strategy when searching for thiol-containing T cell epitopes. Free cysteines can thus be readily transformed into disulphide-like moieties which not only resist undesirable oxidative reactions but which also remain susceptible to reduction by antigen presenting cells, a prerequisite for the activity of thiol-dependent T cell epitopes. We describe the use of this reagent in a study of the intact disulphide-rich protein, toxin alpha from Naja nigricollis, and also two disulphide-containing toxin fragments.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cobra Neurotoxin Proteins / analysis
  • Cysteine / analogs & derivatives*
  • Cysteine / analysis*
  • Cysteine / immunology
  • Cystine / analysis*
  • Cystine / immunology
  • Disulfides / analysis
  • Disulfides / immunology
  • Epitopes / analysis*
  • Evaluation Studies as Topic
  • Lymphocyte Activation
  • Mice
  • Molecular Sequence Data
  • Proteins / analysis*
  • Proteins / immunology
  • Reproducibility of Results
  • Sulfhydryl Compounds / analysis
  • Sulfhydryl Compounds / immunology
  • T-Lymphocytes / immunology*

Substances

  • Cobra Neurotoxin Proteins
  • Disulfides
  • Epitopes
  • Proteins
  • Sulfhydryl Compounds
  • Cystine
  • alpha-cobratoxin
  • t-butyloxycarbonyl-(S-(3-nitro-2-pyridinesulfenyl))cysteine
  • Cysteine