Pseudomonas aeruginosa PilY1 binds integrin in an RGD- and calcium-dependent manner.
Pseudomonas aeruginosa PilY1 binds integrin in an RGD- and calcium-dependent manner.
复制标题
DOI：
10.1371/journal.pone.0029629
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Redinbo MR
中科院分区：
文献类型：
--
作者：
Johnson MD;Garrett CK;Bond JE;Coggan KA;Wolfgang MC;Redinbo MR
PilY 1是来自机会致病菌铜绿假单胞菌的IV型菌毛（tfp）相关蛋白，其与感染性奈瑟氏菌和金氏菌属物种中的相关蛋白具有功能相似性。先前的数据表明，PilY 1作为颤搐运动所必需的钙依赖性菌毛生物发生因子，其特异性钙结合位点位于1，163个残基的蛋白质中的氨基酸850-859处。除了运动性之外，PilY 1还被认为在铜绿假单胞菌tfp与宿主上皮细胞的粘附中起重要作用。在这里，我们表明，PilY 1包含整合素结合甘氨酸-天冬氨酸（RGD）基序位于残基619-621从铜绿假单胞菌的PAK菌株的PilY 1;该基序是保守的从其他已知序列的铜绿假单胞菌菌株的PilY 1。我们证明，纯化的PilY 1结合整合素在体外的RGD依赖性的方式。此外，我们确定了第二个钙结合位点（氨基酸600-608）位于10个残基的上游的RGD。使用D 608 A突变消除该位点的钙结合，消除了整联蛋白结合;相比之下，钙结合模拟物（D 608 K）保留了整联蛋白结合。最后，我们表明，先前建立的PilY 1钙结合位点在851-859也影响蛋白质的协会与整合素。总之，这些数据表明PilY 1在体外以RGD和钙依赖性方式结合整联蛋白。因此，铜绿假单胞菌可以利用这些相互作用来介导体内宿主上皮细胞结合。
PilY1 is a type IV pilus (tfp)-associated protein from the opportunistic pathogen Pseudomonas aeruginosa that shares functional similarity with related proteins in infectious Neisseria and Kingella species. Previous data have shown that PilY1 acts as a calcium-dependent pilus biogenesis factor necessary for twitching motility with a specific calcium binding site located at amino acids 850–859 in the 1,163 residue protein. In addition to motility, PilY1 is also thought to play an important role in the adhesion of P. aeruginosa tfp to host epithelial cells. Here, we show that PilY1 contains an integrin binding arginine-glycine-aspartic acid (RGD) motif located at residues 619–621 in the PilY1 from the PAK strain of P. aeruginosa; this motif is conserved in the PilY1s from the other P. aeruginosa strains of known sequence. We demonstrate that purified PilY1 binds integrin in vitro in an RGD-dependent manner. Furthermore, we identify a second calcium binding site (amino acids 600–608) located ten residues upstream of the RGD. Eliminating calcium binding from this site using a D608A mutation abolished integrin binding; in contrast, a calcium binding mimic (D608K) preserved integrin binding. Finally, we show that the previously established PilY1 calcium binding site at 851–859 also impacts the protein's association with integrin. Taken together, these data indicate that PilY1 binds to integrin in an RGD- and calcium-dependent manner in vitro. As such, P. aeruginosa may employ these interactions to mediate host epithelial cell binding in vivo.
登录
查看更多内容
细胞对血小板传播的附着：Arg-Gly-Asp，钙和整联蛋白受体的作用。
影响因子：
7.8
作者：
Lawler, J;Weinstein, R;Hynes, R O
通讯作者：
Hynes, R O
影响因子：
4
作者：
Humphries JD;Byron A;Humphries MJ
通讯作者：
Humphries MJ
DOI：
10.1016/0006-291x(83)91405-5
发表时间：
1983-01-01
影响因子：
3.1
作者：
AMPHLETT, GW;HRINDA, ME
通讯作者：
HRINDA, ME
影响因子：
5.4
作者：
GOLDMAN, MJ;WILSON, JM
通讯作者：
WILSON, JM
影响因子：
14.8
作者：
Apidianakis, Yiorgos;Rahme, Laurence G.
通讯作者：
Rahme, Laurence G.