[Single-chain, low molecular weight kininogen in the blood plasma of rabbits: isolation and fragmentation by porcine pancreatic kallikrein].
A highly purified preparation of low molecular weight kininogen (LMrK) was isolated from the plasminogen-free rabbit blood plasma, using chromatography on DEAE-Sepharose CL-6B, gel filtration on Ultrogel AcA 34 and Sephadex G-100 as well as gradient chromatography on a hydroxylapatite column. The yield of the 320-fold purified LMrK was 16%. Trypsin released 13-14 micrograms-eq. of bradykinin (BK) from 1 mg of LMrK or 0.85-0,95 mol of BK per mol of kininogen. Rabbit LMrK consists of one polypeptide chain of Mr 69 000 and pI 4.63. Porcine pancreatic kallikrein splits off kinin from the LMrK polypeptide chain by disrupting two peptide bonds resulting in the formation of S-S-bound two chain molecule. After reduction of the S-S bonds by dithioerithritol the latter is separated into a heavy (Mr 61 000) and light (Mr 6 800) chains. A biologically active peptide was isolated from the products of CNBr cleavage of LMrK. This peptide consists of Lys-BK elongated from the C-terminal with several amino acid residues. Rabbit LMrK closely resembles human LMrK in terms of Mr, pI and location of the kinin fragment in the protein molecule.
D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006868:Hydrolysis; D007610:Kallikreins; D007704:Kininogens / Q000097:blood / Q000302:isolation & purification*; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology; D011817:Rabbits; D013552:Swine
D007704:Kininogens; D007610:Kallikreins
Odnotsepochechnyĭ nizkomolekuliarnyĭ kininogen plazmy krovi krolika: vydelenie i kharakter fragmentatsii pankreaticheskim kallikreinom svin'i.