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Functional Characterization of LcpA, a Surface-Exposed Protein of Leptospira spp. That Binds the Human Complement Regulator C4BP

机译:LcpA(钩端螺旋体属表面暴露蛋白)的功能表征。绑定人类补体调节剂C4BP

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We have previously shown that pathogenic leptospiral strains are able to bind C4b binding protein (C4BP). Surface-bound C4BP retains its cofactor activity, indicating that acquisition of this complement regulator may contribute to leptospiral serum resistance. In the present study, the abilities of seven recombinant putative leptospiral outer membrane proteins to interact with C4BP were evaluated. The protein encoded by LIC11947 interacted with this human complement regulator in a dose-dependent manner. The cofactor activity of C4BP bound to immobilized recombinant LIC11947 (rLIC11947) was confirmed by detecting factor I-mediated cleavage of C4b. rLIC11947 was therefore named LcpA (for leptospiral complement regulator-acquiring protein A). LcpA was shown to be an outer membrane protein by using immunoelectron microscopy, cell surface proteolysis, and Triton X-114 fractionation. The gene coding for LcpA is conserved among pathogenic leptospiral strains. This is the first characterization of a Leptospira surface protein that binds to the human complement regulator C4BP in a manner that allows this important regulator to control complement system activation mediated either by the classical pathway or by the lectin pathway. This newly identified protein may play a role in immune evasion by Leptospira spp. and may therefore represent a target for the development of a human vaccine against leptospirosis.
机译:先前我们已经表明,致病性钩端螺旋体菌株能够结合C4b结合蛋白(C4BP)。表面结合的C4BP保留其辅因子活性,表明该补体调节剂的获得可能有助于钩端螺旋体血清抵抗。在本研究中,评估了七个重组推定的钩端螺旋体外膜蛋白与C4BP相互作用的能力。 LIC11947编码的蛋白质以剂量依赖性方式与该人类补体调节剂相互作用。通过检测因子I介导的C4b裂解,可以证实与固定化重组LIC11947(rLIC11947)结合的C4BP的辅因子活性。因此,rLIC11947被命名为LcpA(用于获得钩端螺旋体补体调节剂的蛋白A)。通过使用免疫电子显微镜,细胞表面蛋白水解和Triton X-114分级分离,LcpA被证明是一种外膜蛋白。在病原性钩端螺旋体菌株中,编码LcpA的基因是保守的。这是 Leptospira 表面蛋白的第一个表征,该蛋白以允许该重要调节剂控制经典途径或凝集素途径介导的补体系统激活的方式与人补体调节剂C4BP结合。这种新发现的蛋白质可能在 Leptospira spp的免疫逃避中起作用。因此可能代表开发针对钩端螺旋体病的人类疫苗的目标。

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