Synthesis and immunological evaluation of N-terminal, noncrossreactive tachykinin antigens.
The N-terminal hexa- or pentapeptide sequences of the three mammalian tachykinins substance P, neurokinin A, and neurokinin B have been synthesized by the conventional solid-phase procedure with 6-aminocaproyl-S-(acetamidomethyl)cysteine as a C-terminal spacer and attachment function. A fourth sequence, with an additional N-terminal 6-aminocaproyl residue on the substance P-hapten sequence, was cyclized N- to C-terminally. For this purpose, a four-level protection scheme has been applied: BOC-TFA for N-terminal protection and cleavage; TFA-stable but HF-labile anchoring function and side-chain protection; S-acetamidomethyl for semipermanent thiol protection. The side chain amino function of Lys was protected with NO2Z, stable against HF but readily cleaved with hydrogenation. The hapten sequences were coupled to maleimidated BSA, after the Acm group was removed by mercury/hydrogen sulfide treatment. Mice immunized with the three linear hapten sequences produced sera that were specific in enzyme-linked immunosorbant assay for the presented hapten and the respective tachykinin but displayed no crossreactivity at all toward the other haptens nor to one of the other tachykinins. It is concluded that this approach produced antisera, specific and selective for its respective mammalian tachykinins.
Neugebauer
W
W
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
Elliott
P
P
Cuello
A C
AC
Escher
E
E
D000818:Animals; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D015288:Neurokinin A / Q000276:immunology; D015287:Neurokinin B / Q000276:immunology; D013373:Substance P / Q000276:immunology; D015320:Tachykinins / Q000138:chemical synthesis / Q000276:immunology*
D015320:Tachykinins; D013373:Substance P; D015288:Neurokinin A; D015287:Neurokinin B