Schmidt W, Schmidt M A
Zentrum für Molekulare Biologie, Universität Heidelberg, Federal Republic of Germany.
Infect Immun. 1989 Feb;57(2):438-45. doi: 10.1128/iai.57.2.438-445.1989.
The linear immunogenic and antigenic structure of the S2 subunit of pertussis toxin was investigated with synthetic peptides corresponding to regions of the protein sequence predicted to contain surface-exposed hydrophilic beta turns. Five peptides as peptide-bovine serum albumin conjugates were recognized by anti-pertussis toxin antiserum and were thus designated "immunogenic epitopes." Two prominent immunogenic epitopes were specified by peptides corresponding to sequences spanning R107-120 and R186-199, whereas peptides corresponding to residues R35-50 and R91-106 were only bound in low titer. Three peptides as thyroglobulin conjugates elicited antisera in rabbits that bound intact pertussis toxin by enzyme-linked immunosorbent assay and immunoblot. These peptides were designated "antigenic epitopes." The most prominent antigenic determinant was localized to the N-terminal end of the S2 sequence encompassing residue R1-7. Peptides R35-50 and R91-106 represented two minor antigenic epitopes. Antisera to two additional peptides corresponding to residues R134-149 and R186-199 recognized the S2 subunit only by Western blotting (immunoblotting). Only antiserum raised against peptide R91-106 also recognized the S3 subunit by Western blotting, indicating a marked antigenic and probably also structural difference between the two highly homologous subunits.
利用与蛋白质序列中预测含有表面暴露亲水性β转角区域相对应的合成肽，研究了百日咳毒素S2亚基的线性免疫原性和抗原结构。五种作为肽-牛血清白蛋白偶联物的肽被抗百日咳毒素抗血清识别，因此被指定为“免疫原性表位”。对应于跨越R107 - 120和R186 - 199序列的肽确定了两个突出的免疫原性表位，而对应于R35 - 50和R91 - 106残基的肽仅以低滴度结合。三种作为甲状腺球蛋白偶联物的肽在兔体内引发了抗血清，这些抗血清通过酶联免疫吸附测定和免疫印迹法与完整的百日咳毒素结合。这些肽被指定为“抗原性表位”。最突出的抗原决定簇定位于S2序列的N末端，包括R1 - 7残基。肽R35 - 50和R91 - 106代表两个次要的抗原性表位。针对对应于R134 - 149和R186 - 199残基的另外两种肽产生的抗血清仅通过蛋白质印迹法（免疫印迹法）识别S2亚基。仅针对肽R91 - 106产生的抗血清也通过蛋白质印迹法识别S3亚基，表明这两个高度同源的亚基之间存在明显的抗原性差异，可能也存在结构差异。
