The Haemophilus influenzae Hap autotransporter mediates microcolony formation and adherence to epithelial cells and extracellular matrix via binding regions in the C-terminal end of the passenger domain

被引:58
作者
Fink, DL
Buscher, AZ
Green, B
Fernsten, P
St Geme, JW [1 ]
机构
[1] Washington Univ, Sch Med, Edward Mallinckrodt Dept Pediat, St Louis, MO 63110 USA
[2] Washington Univ, Sch Med, Dept Mol Microbiol, St Louis, MO 63110 USA
[3] St Louis Childrens Hosp, Div Infect Dis, St Louis, MO 63110 USA
[4] Wyeth Vaccines, W Henrietta, NY 14586 USA
关键词
D O I
10.1046/j.1462-5822.2003.00266.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The pathogenesis of non-typable Haemophilus influenzae disease begins with colonization of the nasopharynx and is facilitated by bacterial adherence to respiratory mucosa. The H. influenzae Hap autotransporter is a non-pilus adhesin that promotes adherence to epithelial cells and selected extracellular matrix proteins and mediates bacterial aggregation and microcolony formation. In addition, Hap has serine protease activity. Hap contains a 110 kDa internal passenger domain called Hap(S) and a 45 kDa C-terminal translocator domain called Hapbeta. In the present study, we sought to define the structural basis for Hap adhesive activities. Based on experiments using a panel of monoclonal antibodies against Hap(S) , a deletion derivative lacking most of Hap(S) and a purified fragment of Hap(S) , we established that adherence to epithelial cells is mediated by sequences within the C-terminal 311 residues of Hap(S). In additional experiments, we discovered that bacterial aggregation is also mediated by sequences within the C-terminal 311 residues of Hap(S) and occurs via Hap(S)-Hap(S) interaction between molecules on neighbouring organisms. Finally, we found that adherence to fibronectin, laminin and collagen IV is mediated in part by sequences within the C-terminal 311 residues of Hap(S) and in full by sequences within the C-terminal 511 residues of Hap(S). Taken together, these results demonstrate that all Hap adhesive activities reside in the C-terminal portion of Hap(S). Coupled with earlier observations, the current results establish that Hap(S) adhesive activities and Hap(S) protease activity are contained in separate modules of the protein.
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页码:175 / 186
页数:12
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