Canine plasma kininogen: evidence for the presence of two kininogens and purification of high molecular weight kininogen and characterization as multi-functional molecule.
The ratio of kininogen that is substrate of plasma kallikrein to kininogen, which is not substrate of plasma kallikrein in canine plasma, was about 1:3.6 by differential assay of kininogens. When the plasma was gel-filtered through a column of Sephacryl S-300 superfine, two fractions, which released kinin by trypsin, were obtained. These results indicate that two kininogens with different molecular weights are present in the plasma and they show different susceptibility to plasma kallikrein. One kininogen was purified by ion-exchange and zinc-chelating affinity chromatographies. Purified kininogen showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing condition and its molecular weight was 125 kDa. Released kinin from the kininogen by trypsin was bradykinin. The kininogen inhibited papain and ficin but did not inhibit bromelain at the concentration used. The kininogen bound to carboxymethylated-papain and this binding was dissociated by 3M NaSCN. Canine plasma shortened the abnormal clotting time of human high molecular weight kininogen-deficint plasma. The kininogen also shortened the abnormal clotting time of the plasma. From these results, the purified kininogen was high molecular weight kininogen and it was multi-functional protein.
Mashiko
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H
Division of Chemistry of Hygiene, Meiu College of Pharmacy, Tokyo, Japan.
Takahashi
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H
D000818:Animals; D001780:Blood Coagulation Tests; D002853:Chromatography, Liquid / Q000379:methods; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007610:Kallikreins / Q000378:metabolism; D007704:Kininogens / Q000097:blood* / Q000737:chemistry* / Q000494:pharmacology; D007705:Kinins / Q000378:metabolism; D008970:Molecular Weight; D010206:Papain / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism
D015853:Cysteine Proteinase Inhibitors; D007704:Kininogens; D007705:Kinins; D011498:Protein Precursors; D007610:Kallikreins; D010206:Papain