Biotinylation of protein complexes may lead to aggregation as well as to loss of subunits as revealed by Blue Native PAGE

J Immunol Methods. 2001 Jun 1;252(1-2):171-4. doi: 10.1016/s0022-1759(01)00357-x.

Abstract

Blue Native polyacrylamide gel electrophoresis (BN-PAGE) is a high-resolution method for studying native protein complexes. Here, the migration behaviour of the B cell antigen receptor (BCR) in BN-PAGE with and without prior biotinylation of the cell surface, from which the complexes were purified, are compared. Non-biotinylated complexes appear as a single band, whereas biotinylated complexes display several bands, indicating that biotinylation leads to aggregation of complexes as well as to loss of subunits. Thus, BN-PAGE has limitations in studying multiprotein complexes. These results are used to gain insight into the stoichiometry of the complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin / chemistry*
  • Biotinylation
  • Electrophoresis, Polyacrylamide Gel / methods
  • Immunoglobulin M / chemistry*
  • Receptors, Antigen, B-Cell / chemistry*

Substances

  • Immunoglobulin M
  • Receptors, Antigen, B-Cell
  • Biotin