Purification and characterization of alpha 2-, alpha 2-beta- and beta-macroglobulin inhibitors in the hedgehog, Erinaceus europaeus: beta-macroglobulin identified as the plasma antihemorrhagic factor.
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.
D000818:Animals; D001798:Blood Proteins / Q000032:analysis / Q000302:isolation & purification*; D003429:Cross Reactions; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006364:Hedgehogs / Q000097:blood*; D005779:Immunodiffusion; D008259:Macroglobulins / Q000037:antagonists & inhibitors*; D010206:Papain / Q000037:antagonists & inhibitors; D011480:Protease Inhibitors / Q000302:isolation & purification; D011817:Rabbits
D001798:Blood Proteins; D008259:Macroglobulins; D011480:Protease Inhibitors; C003941:antihemorrhagic factor; D010206:Papain