Altieri D C, Agbanyo F R, Plescia J, Ginsberg M H, Edgington T S, Plow E F
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
J Biol Chem. 1990 Jul 25;265(21):12119-22.
Mac-1 (CD11b/CD18), a leukocyte-restricted integrin receptor, mediates neutrophil/monocyte adhesion to vascular endothelium and phagocytosis of complement-opsonized particles. Recent studies have shown that Mac-1 also functions as a receptor for fibrinogen in a reaction linked to fibrin deposition on the monocyte surface. In this study, we have used extended proteolytic digestion of fibrinogen to identify the region of this molecule that interacts with Mac-1. We found that an Mr approximately 30,000 plasmic fragment D of fibrinogen (D30) produced dose-dependent inhibition (IC50 = 1.6 microM) of the interaction of intact 125I-fibrinogen with stimulated neutrophils and monocytes. 125I-D30 bound saturably to these cells with specific association of 136,200 +/- 15,000 molecules/cell in a reaction inhibited by OKM1 and M1/70, monoclonal antibodies specific for the alpha subunit of Mac-1. Direct microsequence analysis and an epitope-mapped monoclonal antibody showed that D30 lacks the COOH-terminal dodecapeptide of the gamma chain as well as the Arg-Gly-Asp sequences in the A alpha chain. We conclude that fibrinogen interacts with the leukocyte integrin Mac-1 through a novel recognition site that is not shared with other known integrins that function as fibrinogen receptors.
Mac-1(CD11b/CD18)是一种白细胞限制性整合素受体,介导中性粒细胞/单核细胞与血管内皮的黏附以及对补体调理素化颗粒的吞噬作用。最近的研究表明,在与纤维蛋白沉积于单核细胞表面相关的反应中,Mac-1还作为纤维蛋白原的受体发挥作用。在本研究中,我们通过对纤维蛋白原进行延长的蛋白酶消化,以鉴定该分子中与Mac-1相互作用的区域。我们发现,纤维蛋白原的一个分子量约为30,000的血浆片段D(D30)对完整的125I-纤维蛋白原与活化的中性粒细胞和单核细胞之间的相互作用产生剂量依赖性抑制(IC50 = 1.6 microM)。125I-D30以136,200 +/- 15,000分子/细胞的特异性结合量饱和结合这些细胞,该反应可被OKM1和M1/70抑制,这两种单克隆抗体对Mac-1的α亚基具有特异性。直接的微序列分析和一种表位定位的单克隆抗体表明,D30缺乏γ链的COOH末端十二肽以及Aα链中的Arg-Gly-Asp序列。我们得出结论,纤维蛋白原通过一个新的识别位点与白细胞整合素Mac-1相互作用,该识别位点与其他作为纤维蛋白原受体的已知整合素不共享。