Helicobacter pylori VacA subdomain required for intracellular toxin activity and assembly of functional oligomeric complexes

被引:32
作者
Ivie, Susan E.
McClain, Mark S. [2 ]
Torres, Victor J.
Algood, Holly M. Scott [2 ]
Lacy, D. Borden
Yang, Rong [4 ,5 ]
Blanke, Steven R. [4 ,5 ]
Cover, Timothy L. [1 ,2 ,3 ]
机构
[1] Vanderbilt Univ, Sch Med, Div Infect Dis, Dept Microbiol & Immunol, Nashville, TN 37232 USA
[2] Vanderbilt Univ, Sch Med, Dept Med, Nashville, TN 37232 USA
[3] Vet Affairs Tennessee Valley Healthcare Syst, Nashville, TN 37212 USA
[4] Univ Illinois, Dept Microbiol, Urbana, IL 61801 USA
[5] Univ Illinois, Inst Genom Biol, Urbana, IL 61801 USA
关键词
D O I
10.1128/IAI.01664-07
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Helicobacter pylori VacA is a secreted pore-forming toxin that is comprised of two domains, designated p33 and p55. The p55 domain has an important role in the binding of VacA to eukaryotic cell surfaces. A total of 111 residues at the amino terminus of p55 (residues 312 to 422) are essential for the intracellular activity of VacA, which suggests that this region may constitute a subdomain with an activity distinct from cell binding. To investigate the properties of this subdomain, a small deletion mutation (targeting aspartic acid 346 and glycine 347) was introduced into the H. pylori chromosomal vacA gene. Similar to wild-type VacA, the VacA Delta 346-347 mutant protein was proteolytically processed, secreted, and bound to eukaryotic cells. However, VacA Delta 346-347 did not cause cell vacuolation or membrane depolarization, and it was impaired in the ability to assemble into large water-soluble oligomeric structures. Interestingly, VacA Delta 346-347 was able to physically interact with wild-type VacA to form mixed oligomeric complexes, and VacA Delta 346-347 inhibited wild-type vacuolating activity in a dominant-negative manner. These data indicate that the assembly of functional oligomeric VacA complexes is dependent on specific sequences, including amino acids 346 and 347, within the p55 amino-terminal subdomain.
引用
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页码:2843 / 2851
页数:9
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