Isolation and characterization of a conserved porin protein from Helicobacter pylori.
Helicobacter pylori is a causative agent of gastritis in humans and is correlated with gastric ulcer formation. Infections with this bacterium have proven difficult to treat with antimicrobial agents. To better understand how this bacterium transports compounds such as antimicrobial agents across its outer membrane, identification of porin proteins is important. We have recently identified a family of H. pylori porins (HopA to HopD) (M. M. Exner, P. Doig, T. J. Trust, and R. E. W. Hancock, Infect. Immun. 63:1567-1572, 1995). Here, we report on an unrelated porin species (HopE) from this bacterium. This protein had a apparent molecular mass of 31 kDa and was seen to form 50- and 90-kDa aggregates that were designated putative dimeric and trimeric forms, respectively. The protein was purified to homogeneity and, with a model planar lipid membrane system, was shown to act as a nonselective pore with a single channel conductance in 1.0 M KCl of 1.5 nS, similarly to other bacterial nonspecific porins. An internal peptide sequence of HopE shared homology with the P2 porin of Haemophilus influenzae. HopE was also shown to be antigenic in vivo as assessed by sera taken from H. pylori-infected individuals and was immunologically conserved with both patient sera and specific monoclonal antibodies. From these data, it appears that HopE is a major nonselective porin of H. pylori. The implications of these findings are discussed.
Doig
P
P
Canadian Bacterial Diseases Network, University of Victoria, British Columbia, Canada.
Exner
M M
MM
Hancock
R E
RE
Trust
T J
TJ
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D004553:Electric Conductivity; D016480:Helicobacter pylori / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D008051:Lipid Bilayers; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D018272:Porins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013388:Succinimides
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D008051:Lipid Bilayers; D010446:Peptide Fragments; D018272:Porins; D013388:Succinimides; C011240:dithiobis(succinimidylpropionate)