Complement Factor H-Related Proteins CFHR2 and CFHR5 Represent Novel Ligands for the Infection-Associated CRASP Proteins of Borrelia burgdorferi
Complement Factor H-Related Proteins CFHR2 and CFHR5 Represent Novel Ligands for the Infection-Associated CRASP Proteins of Borrelia burgdorferi
复制标题
补体因子 H 相关蛋白 CFHR2 和 CFHR5 代表伯氏疏螺旋体感染相关 CRASP 蛋白的新型配体
作者：
C. Siegel;T. Hallström;C. Skerka;H. Eberhardt;B. Uzonyi;Tobias Beckhaus;M. Karas;R. Wallich;B. Stevenson;P. Zipfel;P. Kraiczy
伯氏疏螺旋体的毒力特性之一是其对先天免疫的抗性，特别是对补体介导的杀伤。抗血清B。Burgdorferi表达多达五种不同的补体调节获得表面蛋白（CRASP），其与补体调节因子H（CFH）和因子H样蛋白1（FHL 1）或因子H相关蛋白1（CFHR 1）相互作用。在本研究中，我们阐明了感染相关的CRASP-3和CRASP-5蛋白作为额外的补体调节蛋白的配体以及B的补体抵抗的作用。burgdorferi。为了阐明CRASP-5和CRASP-3是否与各种人蛋白质相互作用，将两种疏螺旋体蛋白质固定在磁珠上。与人血清孵育后，洗脱结合的蛋白质并通过甘氨酸-SDS-PAGE分离。除了CFH和CFHR 1，补体调节因子CFHR 2和CFHR 5被确定为新的配体的疏螺旋体蛋白采用MALDI-TOF。为了进一步评估CRASP-3和CRASP-5对补体抗性的贡献，使用血清敏感的B.将缺乏所有CFH结合蛋白的伽氏菌菌株G1用作功能分析的有价值的模型。两种CRASP均在B上表达。ELISA结果显示，与CFH、CFHR 1和CFHR 2结合的细胞数均高于与CFH、CFHR 1和CFHR 2的结合。相比之下，直播B.血清吸附试验和流式细胞仪分析表明，Garinii结合CFHR 1、CFHR 2和CFHR 5，仅结合极少量的CFH。进一步的功能分析显示，在NHS孵育后，表达CRASP-3或CRASP-5的疏螺旋体被补体杀死。结论/意义在CFH不存在和CFHR 1、CFHR 2和CFHR 5存在的情况下，膜攻击复合物的组装和整合没有被有效抑制，表明CFH与CFHR 1、CFHR 2和CFHR 5协同作用支持B的补体逃避。burgdorferi。
Background One virulence property of Borrelia burgdorferi is its resistance to innate immunity, in particular to complement-mediated killing. Serum-resistant B. burgdorferi express up to five distinct complement regulator-acquiring surface proteins (CRASP) which interact with complement regulator factor H (CFH) and factor H-like protein 1 (FHL1) or factor H-related protein 1 (CFHR1). In the present study we elucidate the role of the infection-associated CRASP-3 and CRASP-5 protein to serve as ligands for additional complement regulatory proteins as well as for complement resistance of B. burgdorferi. Methodology/Principal Findings To elucidate whether CRASP-5 and CRASP-3 interact with various human proteins, both borrelial proteins were immobilized on magnetic beads. Following incubation with human serum, bound proteins were eluted and separated by Glycine-SDS-PAGE. In addition to CFH and CFHR1, complement regulators CFHR2 and CFHR5 were identified as novel ligands for both borrelial proteins by employing MALDI-TOF. To further assess the contributions of CRASP-3 and CRASP-5 to complement resistance, a serum-sensitive B. garinii strain G1 which lacks all CFH-binding proteins was used as a valuable model for functional analyses. Both CRASPs expressed on the B. garinii outer surface bound CFH as well as CFHR1 and CFHR2 in ELISA. In contrast, live B. garinii bound CFHR1, CFHR2, and CFHR5 and only miniscute amounts of CFH as demonstrated by serum adsorption assays and FACS analyses. Further functional analysis revealed that upon NHS incubation, CRASP-3 or CRASP-5 expressing borreliae were killed by complement. Conclusions/Significance In the absence of CFH and the presence of CFHR1, CFHR2 and CFHR5, assembly and integration of the membrane attack complex was not efficiently inhibited indicating that CFH in co-operation with CFHR1, CFHR2 and CFHR5 supports complement evasion of B. burgdorferi.
影响因子：
20.3
作者：
Heinen, Stefan;Hartmann, Andrea;Skerka, Christine
通讯作者：
Skerka, Christine
纤维蛋白原是一种新型脂蛋白颗粒的组成部分：H 因子相关蛋白 (FHRP) 相关脂蛋白颗粒 (FALP)。
影响因子：
20.3
作者：
Park,CT;Wright,SD
通讯作者：
Wright,SD