Worobec E A, Taneja A K, Hodges R S, Paranchych W
J Bacteriol. 1983 Feb;153(2):955-61. doi: 10.1128/jb.153.2.955-961.1983.
Trypsin digestion of pilin monomers from EDP208 conjugative pili causes cleavage of Lys12 to yield an N-terminal dodecapeptide, ET1 (Mr approximately equal to 1,500), and the remaining C-terminal fragment, ER (Mr approximately equal to 10,000). Using the amino acid sequence for ET1 provided by Frost et al. (J. Bacteriol. 153:950-954), we synthesized the N-terminal dodecapeptide chemically, conjugated it to bovine serum albumin, and subjected it to immunological studies. Antisera prepared against intact EDP208 pili as well as against the synthetic ET1-BSA conjugate were used in experiments involving an enzyme-linked immunosorbant assay and electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to nitrocellulose sheets. Both experimental approaches showed strong reactivity between the synthetic dodecapeptide and antiserum raised against whole pili. It was also found that antiserum raised against the synthetic peptide was reactive against intact pilus protein, indicating that the N-terminal dodecapeptide is an important antigenic determinant of the EDP208 pilus protein. Additional studies showed that the C-terminal fragment, ER, may contain one or two additional antigenic sites.
用胰蛋白酶消化来自EDP208接合菌毛的菌毛单体，会使赖氨酸12发生裂解，产生一个N端十二肽ET1（分子量约为1500）和剩余的C端片段ER（分子量约为10000）。利用弗罗斯特等人提供的ET1氨基酸序列（《细菌学杂志》153:950 - 954），我们化学合成了N端十二肽，将其与牛血清白蛋白偶联，并进行免疫学研究。针对完整的EDP208菌毛以及合成的ET1 - BSA偶联物制备的抗血清，用于涉及酶联免疫吸附测定以及蛋白质从十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳转移至硝酸纤维素膜的实验。两种实验方法均显示合成的十二肽与针对完整菌毛产生的抗血清之间有强烈反应性。还发现针对合成肽产生的抗血清与完整菌毛蛋白有反应，这表明N端十二肽是EDP208菌毛蛋白的一个重要抗原决定簇。进一步研究表明，C端片段ER可能包含一个或两个额外的抗原位点。
