Tschopp J, Masson D, Schäfer S, Peitsch M, Preissner K T
Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland.
Biochemistry. 1988 May 31;27(11):4103-9. doi: 10.1021/bi00411a029.
S-Protein/vitronectin is a serum glycoprotein that inhibits the lytic activity of the membrane attack complex of complement, i.e., of the complex including the proteins C5b, C6, C7, C8, and C9n. We show that intact S-protein/vitronectin or its cyanogen bromide generated fragments also inhibit the hemolysis mediated by perforin from cytotoxic T-cells at 45 and 11 microM, respectively. The glycosaminoglycan binding site of S-protein/vitronectin is responsible for the inhibition, since a synthetic peptide corresponding to a part of this highly basic domain (amino acid residues 348-360) inhibits complement- as well as perforin-mediated cytolysis. In the case of C9, the synthetic peptide binds to the acidic residues occurring in its N-terminal cysteine-rich domain (residues 101-111). Antibodies raised against this particular segment react 25-fold better with the polymerized form of C9 as compared with its monomeric form, indicating that this site becomes exposed only upon the hydrophilic-amphiphilic transition of C9. Since the cysteine-rich domain of C9 has been shown to be highly conserved in C6, C7, and C8 as well as in perforin, the inhibition of the lytic activities of these molecules by S-protein/vitronectin or by peptides corresponding to its heparin binding site may be explained by a similar mechanism.
S-蛋白/玻连蛋白是一种血清糖蛋白,可抑制补体膜攻击复合物的溶解活性,即包括蛋白质C5b、C6、C7、C8和C9n的复合物的溶解活性。我们发现,完整的S-蛋白/玻连蛋白或其溴化氰产生的片段也分别在45微摩尔和11微摩尔时抑制细胞毒性T细胞穿孔素介导的溶血。S-蛋白/玻连蛋白的糖胺聚糖结合位点负责这种抑制作用,因为对应于这个高度碱性结构域一部分(氨基酸残基348 - 360)的合成肽可抑制补体以及穿孔素介导的细胞溶解。就C9而言,合成肽与C9 N端富含半胱氨酸结构域(残基101 - 111)中出现的酸性残基结合。针对这一特定片段产生的抗体与聚合形式的C9反应比与单体形式的C9反应好25倍,表明该位点仅在C9发生亲水 - 亲脂转变时才暴露。由于已证明C9富含半胱氨酸的结构域在C6、C7和C8以及穿孔素中高度保守,S-蛋白/玻连蛋白或与其肝素结合位点对应的肽对这些分子溶解活性的抑制作用可能由类似机制解释。