Two-dimensional blue native/SDS gel electrophoresis of multiprotein complexes from Helicobacter pylori
Molecular & Cellular Proteomics. 2007-02-01; 6(2): 193-206
DOI: 10.1074/mcp.M600363-MCP200
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1. Mol Cell Proteomics. 2007 Feb;6(2):193-206. Epub 2006 Nov 7.
Two-dimensional blue native/SDS gel electrophoresis of multiprotein complexes
from Helicobacter pylori.
Pyndiah S(1), Lasserre JP, Ménard A, Claverol S, Prouzet-Mauléon V, Mégraud F,
Zerbib F, Bonneu M.
Author information:
(1)INSERM, U853, Bordeaux, F 33076 France.
The study of protein interactions constitutes an important domain to understand
the physiology and pathogenesis of microorganisms. The two-dimensional blue
native/SDS-PAGE was initially reported to analyze membrane protein complexes. In
this study, both cytoplasmic and membrane complexes of a bacterium, the strain
J99 of the gastric pathogen Helicobacter pylori, were analyzed by this method. It
was possible to identify 34 different proteins grouped in 13 multiprotein
complexes, 11 from the cytoplasm and two from the membrane, either previously
reported partially or totally in the literature. Besides complexes involved in H.
pylori physiology, this method allowed the description of interactions involving
known pathogenic factors such as (i) urease with the heat shock protein GroEL or
with the putative ketol-acid reductoisomerase IlvC and (ii) the cag pathogenicity
island CagA protein with the DNA gyrase GyrA as well as insight on the partners
of TsaA, a peroxide reductase/stress-dependent molecular chaperone. The
two-dimensional blue native/SDS-PAGE combined with mass spectrometry is a
potential tool to study the differences in complexes isolated in various
situations and also to study the interactions between bacterial and eucaryotic
cell proteins.
DOI: 10.1074/mcp.M600363-MCP200
PMID: 17092930 [Indexed for MEDLINE]