de Wit C A, Weström B R
Department of Zoophysiology, University of Lund, Sweden.
Toxicon. 1987;25(11):1209-19. doi: 10.1016/0041-0101(87)90139-5.
Three macroglobulin inhibitors were purified from hedgehog (Erinaceus europaeus) plasma by sequential chromatography on Cibacron Blue Sepharose, Sephacryl S-200 and preparative agarose gel electrophoresis. Each macroglobulin was characterized for proteinase inhibiting activity, molecular weight by polyacrylamide gel electrophoresis (PAGE), subunit size by sodium dodecyl sulfate (SDS)-PAGE, immunological cross-reactivity to other macroglobulins and antihemorrhagic activity against European viper (Vipera berus) venom. Hedgehog alpha 2-macroglobulin is a tetramer (Mr 800,000) composed of identical monomers (Mr 200,000) that inhibits all proteinases tested and is the homologue of human alpha 2-macroglobulin, rat alpha 2-acute phase globulin, dog alpha 1-macroglobulin and swine alpha 2-macroglobulin fast. Hedgehog alpha 2-beta-macroglobulin is a dimer (Mr 450-550,000) composed of identical monomers (Mr 200,000) that inhibits all proteinases tested and appears to be structurally similar to other animal 'half-molecule' macroglobulins. Hedgehog beta-macroglobulin (Mr 700,000) gave subunits of 34,000 and 39,000 after SDS-PAGE and showed cross-reactivity with swine alpha 2-macroglobulin slow. It inhibits all proteinases tested and is the only macroglobulin with antihemorrhagic activity against V. berus venom. This antihemorrhagic activity may be due to beta-macroglobulin's different structure as compared to other macroglobulins, which may make it less susceptible to inactivation by venom proteinases.
通过在Cibacron Blue Sepharose、Sephacryl S - 200上的连续色谱法以及制备性琼脂糖凝胶电泳，从刺猬（欧洲刺猬）血浆中纯化出三种巨球蛋白抑制剂。对每种巨球蛋白进行了蛋白酶抑制活性、聚丙烯酰胺凝胶电泳（PAGE）测定的分子量、十二烷基硫酸钠（SDS）-PAGE测定的亚基大小、与其他巨球蛋白的免疫交叉反应性以及对欧洲蝰蛇（极北蝰）毒液的抗出血活性等特性分析。刺猬α2 - 巨球蛋白是一种四聚体（分子量800,000），由相同的单体（分子量200,000）组成，可抑制所有测试的蛋白酶，是人类α2 - 巨球蛋白、大鼠α2 - 急性期球蛋白、犬α1 - 巨球蛋白和猪α2 - 巨球蛋白快速型的同源物。刺猬α2 - β - 巨球蛋白是一种二聚体（分子量450 - 550,000），由相同的单体（分子量200,000）组成，可抑制所有测试的蛋白酶，并且在结构上似乎与其他动物的“半分子”巨球蛋白相似。刺猬β - 巨球蛋白（分子量700,000）在SDS - PAGE后产生分子量为34,000和39,000的亚基，并与猪α2 - 巨球蛋白慢速型显示交叉反应性。它可抑制所有测试的蛋白酶，是唯一对极北蝰毒液具有抗出血活性的巨球蛋白。这种抗出血活性可能是由于β - 巨球蛋白与其他巨球蛋白相比结构不同，这可能使其较不易被毒液蛋白酶灭活。
