CHARACTERIZATION OF THE PSEUDOMONAS-AERUGINOSA PILUS ADHESIN - CONFIRMATION THAT THE PILIN STRUCTURAL PROTEIN SUBUNIT CONTAINS A HUMAN EPITHELIAL CELL-BINDING DOMAIN
CHARACTERIZATION OF THE PSEUDOMONAS-AERUGINOSA PILUS ADHESIN - CONFIRMATION THAT THE PILIN STRUCTURAL PROTEIN SUBUNIT CONTAINS A HUMAN EPITHELIAL CELL-BINDING DOMAIN
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DOI：
10.1128/iai.57.12.3720-3726.1989
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发表时间：
1989-12-01
影响因子：
3.1
通讯作者：
HODGES, RS
中科院分区：
文献类型：
--
作者：
IRVIN, RT;DOIG, P;HODGES, RS
先前的研究表明，铜绿假单胞菌 PAK 菌毛粘附素部分存在于位于 PAK 菌毛蛋白结构蛋白 C 末端区域的上皮细胞结合域中。合成肽 Ac17red（与 PAK 菌毛蛋白残基 128 至 144 序列相同的合成肽，其中 Cys-129 和 Cys-142 残基处于还原状态）和 Ac17ox（与 PAK 菌毛蛋白残基 128 至 144 序列相同的合成肽，在氨基酸残基 Cys-129 和 Cys-129 之间形成二硫键合成了应包含上皮细胞结合结构域的 Cys-142)。 Ac17red 和 Ac17ox 均与颊上皮细胞 (BEC) 和纤毛气管上皮细胞 (TEC) 结合。 Ac17ox 与 BEC 的结合 Km 为 6.40 μM，而 Ac17red 的 Km 为 9.87 μM。 Ac17red 与纯化菌毛所结合的受体位点相同，并竞争性抑制纯化 PAK 菌毛与 BEC 的结合。固定在硝化纤维素上的分子量为 82、55 至 51 和 40 千道尔顿的 BEC 糖蛋白对 Ac17red 表现出高碘酸盐敏感受体活性； PAK pili 也有类似的活性。 Ac17red、Ac17ox 和 PAK 菌毛与人 TEC 细胞质膜的纤毛和管腔部分结合，与铜绿假单胞菌全细胞结合的区域相同。 PAK 菌毛蛋白具有位于蛋白质 C 末端区域的上皮细胞结合结构域。
Previous studies have suggested that the Pseudomonas aeruginosa PAK pilus adhesin moiety resides in an epithelial cell-binding domain located in the C-terminal region of the PAK pilin structural protein. Synthetic peptides Ac17red (a synthetic peptide with a sequence identical to that of PAK pilin residues 128 to 144, with the Cys-129 and Cys-142 residues being in the reduced state) and Ac17ox (a synthetic peptide with a sequence identical to that of PAK pilin residues 128 to 144, with a formed disulfide bridge beween the amino acid residues Cys-129 and Cys-142), which should contain the epithelial cell-binding domain, were synthesized. Ac17red and Ac17ox both bound to buccal epithelial cells (BECs) and to cilitated tracheal epithelial cells (TECs). Ac17ox had a Km of 6.40 .mu.M for binding to BECs, while Ac17red had a Km of 9.87 .mu.M. Ac17red bound to the same receptor sites that purified pili did and competitively inhibited the binding of purified PAK pili to BECs. BEC glycoproteins with molecular masses of 82, 55 to 51, and 40 kilodaltons immobilized on nitrocellulose exhibited periodate-sensitive receptor activity for Ac17red; similar activity has been found for PAK pili. Ac17red, Ac17ox, and PAK pili bound to the cilia and luminal portions of the cytoplasmic membrane of human TECs, the same regions to which P. aeruginosa whole cells bind. PAK pilin has an epithelial cell-binding domain that resides in the C-terminal region of the protein.