Boc-Cys(Npys)-OH (BCNP): an appropriate reagent for the identification of T cell epitopes in cystine and/or cysteine-containing proteins.
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.
Mourier
G
G
Département d'Ingénierie et d'Etudes des Protéines, C.E.A., Bt 152, C.E. Saclay, Gif-sur-Yvette, France.
Maillère
B
B
Cotton
J
J
Hervé
M
M
Leroy
S
S
Léonetti
M
M
Ménez
A
A
D000595:Amino Acid Sequence; D000818:Animals; D003039:Cobra Neurotoxin Proteins / Q000032:analysis; D003545:Cysteine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000276:immunology; D003553:Cystine / Q000032:analysis* / Q000276:immunology; D004220:Disulfides / Q000032:analysis / Q000276:immunology; D000939:Epitopes / Q000032:analysis*; D005069:Evaluation Studies as Topic; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D015203:Reproducibility of Results; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D003039:Cobra Neurotoxin Proteins; D004220:Disulfides; D000939:Epitopes; D011506:Proteins; D013438:Sulfhydryl Compounds; D003553:Cystine; C025154:alpha-cobratoxin; C050518:t-butyloxycarbonyl-(S-(3-nitro-2-pyridinesulfenyl))cysteine; D003545:Cysteine