Two-dimensional blue native/SDS gel electrophoresis of multiprotein complexes from Helicobacter pylori.
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.
Pyndiah
Slovénie
S
INSERM, U853, Bordeaux, F 33076 France.
Lasserre
Jean Paul
JP
Ménard
Armelle
A
Claverol
Stéphane
S
Prouzet-Mauléon
Valérie
V
Mégraud
Francis
F
Zerbib
Frank
F
Bonneu
Marc
M
D000942:Antigens, Bacterial / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D016480:Helicobacter pylori / Q000737:chemistry* / Q000201:enzymology / Q000472:pathogenicity; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000032:analysis*; D014510:Urease / Q000378:metabolism; D014774:Virulence
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D008565:Membrane Proteins; D046912:Multiprotein Complexes; C080866:cagA protein, Helicobacter pylori; D014510:Urease