CROSS-LINKING OF PORIN WITH GLUTARDIALDEHYDE - A TEST FOR THE ADEQUACY OF PREMISES OF CROSS-LINKING THEORY

被引:8
作者
AZEM, A
SHAKED, I
ROSENBUSCH, JP
DANIEL, E
机构
[1] TEL AVIV UNIV,GEORGE S WISE FAC LIFE SCI,DEPT BIOCHEM,IL-69978 TEL AVIV,ISRAEL
[2] UNIV BASEL,BIOCTR,DEPT MICROBIOL,CH-4056 BASEL,SWITZERLAND
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 1995年 / 1243卷 / 02期
关键词
POLYPEPTIDE CHAIN CROSS-LINKING; CROSS-LINKING THEORY; PORIN; GLUTARDIALDEHYDE; (ESCHERICHIA-COLI);
D O I
10.1016/0304-4165(94)00107-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Exposure of porin from Escherichia coli to glutardialdehyde followed by SDS-gel electrophoresis in 3% polyacrylamide yielded three bands that were identified in order of decreasing mobility as monomers and two and three cross-linked polypeptide chains. The distribution of protein among the three species for different extents of reaction showed a remarkably good agreement with corresponding values predicted from cross linking theory for an oligomer composed of three identical subunits arranged according to a 3-fold rotation axis. Electrophoresis performed in 7.5% polyacrylamide yielded four bands that were assigned to polypeptide chain monomers, dimers, and two types of trimers carrying two and three intersubunit cross-links. Our findings provide evidence that the central premise of cross-linking theory, viz, that the intersubunit cross-links formed upon exposure of a protein to a bifunctional reagent be governed by the symmetry of the molecule, is valid. Careful interpretation of cross-linking experiments thus proves an effective method to assess the oligomeric structure of a protein and reveal the symmetry underlying the spatial arrangement of the subunits within the molecule.
引用
收藏
页码:151 / 156
页数:6
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