Referenced in: none

Automatically associated channels: Kv10.1

Title: Cross-linking of porin with glutardialdehyde: a test for the adequacy of premises of cross-linking theory.

Authors: A Azem, I Shaked, J P Rosenbusch, E Daniel

Journal, date & volume: Biochim. Biophys. Acta, 1995 Feb 23 , 1243, 151-6

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/7873557

Abstract
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.