Levina G O, Makevnina L G, Sherstiuk S F, Paskhina T S
Biokhimiia. 1985 Feb;50(2):325-36.
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.
从不含纤溶酶原的兔血浆中分离出一种高度纯化的低分子量激肽原（LMrK）制剂，采用DEAE-琼脂糖CL-6B柱层析、Ultrogel AcA 34和Sephadex G-100凝胶过滤以及羟基磷灰石柱梯度层析法。320倍纯化的LMrK产率为16%。胰蛋白酶从1mg LMrK中释放出13 - 14微克当量的缓激肽（BK），即每摩尔激肽原释放0.85 - 0.95摩尔BK。兔LMrK由一条分子量为69000、等电点为4.63的多肽链组成。猪胰激肽释放酶通过破坏两个肽键从LMrK多肽链上裂解下激肽，导致形成S-S键连接的双链分子。用二硫苏糖醇还原S-S键后，后者分离为重链（分子量61000）和轻链（分子量6800）。从LMrK的溴化氰裂解产物中分离出一种生物活性肽。该肽由从C末端延伸的Lys-BK和几个氨基酸残基组成。兔LMrK在分子量、等电点以及激肽片段在蛋白质分子中的位置方面与人类LMrK非常相似。
