Premium
Chromatographic Study of Spirosin by Use of Monoclonal Antibodies to Spirosin of Yersinia enterocolitica
Author(s) -
Yamato Masayuki,
Tomotake Hiroyuki,
Shiomoto Yasuhisa,
Ota Fusao
Publication year - 1995
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1995.tb02198.x
Subject(s) - yersinia enterocolitica , monoclonal antibody , biology , enterobacteriaceae , escherichia coli , microbiology and biotechnology , protease , antibody , sepharose , biochemistry , bacteria , enzyme , immunology , genetics , gene
Two monoclonal antibodies (MAbs) reacting with spirosins from Enterobacteriaceae were obtained in a course of screening MAbs to spirosin from Yersinia enterocolitica SYT‐11‐72 (YE72). The antibodies were designated MAbs‐S44 and S50. They were IgG 2b and IgG 2a , respectively, both with k light chains. On Western blotting after limited proteolysis of YE72 spirosin with Staphylococcus aureus V8 protease, they reacted markedly with peptide fragments of 27 and 35 kDa, suggesting the presence of an antigenic determinant on the fragments. When supernatant cell lysate from Escherichia coli K12 was chromatographed on DEAE‐cellulose and Sepharose CL‐6B columns successively, a 96‐kDa protein with alcohol dehydrogenase (ADH) activity was always associated with reactivity to MAb‐S50. These findings combined with N‐terminal amino acid sequences clearly indicate the identity of spirosin to ADH in E. coli .