[Purification and properties of high-molecular-weight rabbit kininogen].
A preparation of high-molecular weight kininogen (HMWK) was isolated from rabbit citrate blood plasma and purified using chromatography on DEAE-Sephadex A-50 and CM-Sephadex C-50. From 1 mg HMWK, trypsin or kallikreine of human blood plasma release 10 mkg bradykinine. The HMWK preparation is homogeneous during electrophoresis in 7.5% polyacrylamide gel in tris-glycine buffer, pH 8.3; its electrophoretic mobility corresponds to that of alpha2-globulins. The molecular weight of HMWK estimated using the collumn with Sephadex G-200, is 130.000--150.000; the sedimentation constant S20w is 7.6. Rabbit HMWK is neither a dimer, nor a trimer of low molecular weight kininogen (LMWK), since it does not degrade into subunits after treatment by 2.5% solution of sodium dodecyl sulfate, containing 8 M urea. 0.05 M 2-mercaptoethanol and 8 M urea induce HMWK splitting into 2 fragments with respective molecular weights of 80.000 and 30.000, the kinine-containing group being localized in the low-molecular weight fragment. Estimation of rates of kinine formation by different kininogenasses from highly purified HMWK and LMWK preparations showed that those kininogens are functionally different substrates, since blood plasma kallikreines release kinines from HMWK at a greater rates, whereas tissue kallikreines, e. g. human saliva kallikreine release kinines from LMWK. The specificity of kallikreines as kininogenase, to trypsin, was determined. Tripsin removes bradykinine from both kininogens at the same rates, which are an order of magnitude less than those found for kallikreines.
D000818:Animals; D001920:Bradykinin / Q000302:isolation & purification; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007610:Kallikreins / Q000097:blood / Q000494:pharmacology; D007704:Kininogens / Q000032:analysis / Q000097:blood / Q000302:isolation & purification*; D007705:Kinins / Q000302:isolation & purification; D008623:Mercaptoethanol / Q000494:pharmacology; D008970:Molecular Weight; D011817:Rabbits; D012463:Saliva / Q000201:enzymology; D014357:Trypsin / Q000032:analysis; D014508:Urea / Q000494:pharmacology
D007704:Kininogens; D007705:Kinins; D008623:Mercaptoethanol; D014508:Urea; D007610:Kallikreins; D014357:Trypsin; D001920:Bradykinin
Ochistka i svoĭstva vysokomolekuliarnogo kininogena krolika.