Bohnsack J F, Mollison K W, Buko A M, Ashworth J C, Hill H R
Department of Pediatrics, University of Utah School of Medicine, Salt Lake City 84132.
Biochem J. 1991 Feb 1;273 ( Pt 3)(Pt 3):635-40. doi: 10.1042/bj2730635.
Incubation of recombinant human C5a (rC5a) with the 7360 strain of group B streptococci (GBS) destroyed the ability of rC5a to stimulate chemotaxis or adherence of purified human polymorphonuclear leucocytes (PMNs). Treatment of 125I-labelled rC5a with GBS 7360 correspondingly decreased rC5a binding to human PMNs. This also resulted in an approx. 600 Da decrease in the molecular mass of rC5a as determined by SDS/PAGE. Incubation of rC5a with the GBS strain GW, which only minimally altered the ability of rC5a to activate human PMNs, did not affect rC5a binding to PMNs and did not alter the molecular mass of rC5a on SDS/PAGE. Plasma-desorption m.s. of rC5a inactivated by GBS 7360 showed that the GBS cleaved the rC5a between histidine-67 and lysine-68 near the C-terminus of rC5a. This mechanism of inactivation of C5a by proteolytic cleavage at the C-terminus of C5a is consistent with the known critical role of the C-terminus in C5a activation of human PMNs. This C5a-cleaving proteinase activity may contribute to the pathophysiology of GBS infections.
将重组人C5a(rC5a)与B族链球菌(GBS)7360菌株共同孵育,会破坏rC5a刺激纯化的人多形核白细胞(PMN)趋化或黏附的能力。用GBS 7360处理125I标记的rC5a,相应地会降低rC5a与人PMN的结合。这也导致通过SDS/PAGE测定的rC5a分子量大约降低600 Da。将rC5a与GBS菌株GW共同孵育,该菌株只会轻微改变rC5a激活人PMN的能力,不会影响rC5a与PMN的结合,也不会改变SDS/PAGE上rC5a的分子量。经GBS 7360灭活的rC5a的等离子体解吸质谱显示,GBS在rC5a C末端附近的组氨酸-67和赖氨酸-68之间切割rC5a。通过在C5a的C末端进行蛋白水解切割来使C5a失活的这种机制,与C末端在人PMN的C5a激活中的已知关键作用一致。这种切割C5a的蛋白酶活性可能有助于GBS感染的病理生理学过程。