Macartney H W, Tschesche H
Eur J Biochem. 1983 Jan 17;130(1):85-92. doi: 10.1111/j.1432-1033.1983.tb07120.x.
A beta 1-serum component, beta 1-anticollagenase, capable of inhibiting various mammalian tissue collagenases, was isolated from human plasma by gel filtration, affinity chromatography and ion-exchange chromatography. The inhibitor contains 1-2 free sulfhydryl groups, which are a prerequiste for inhibitory activity and for binding to the thiol-Sepharose affinity support. Alkylation of beta 1-anticollagenase by iodoacetamide blocks inhibitory activity. The inhibitor was purified to apparent homogeneity and exhibited a Mr = 30500 determined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The amino acid and carbohydrate composition was determined. According to its composition and the isoelectric focussing beta 1-anticollagenase is an acidic protein with an isoelectric point of 5.6. Inhibition of human leukocyte collagenase proceeds in a strong 1 : 1 stoichiometric reaction. The mechanism of this association takes place by a disulfide/thiol interchange reaction as has been previously indicated for human leukocyte collagenases in forming the latent enzyme [Macartney, H. W. and Tschesche, H. (1980) FEBS Lett. 119, 327-332]. The beta 1-anticollagenase--leukocyte-collagenase complex (latent enzyme) is activatable by disulfide-containing compounds such as cystine, oxidised glutathione, insulin, relaxin, trypsinogen and others, but not by 179,203-di(S-carboxymethyl)trypsinogen, or its trypsin derivative. Compounds containing inaccessible disulfide bonds, e.g. chymotrypsin, or sulfhydryl groups, e.g. D-penicillamine, do not activate the complex. Activation is, however, easily obtained with the oxidised-glutathione-generating system myeloperoxidase/H2O2/glutathione as was previously demonstrated for the human leukocyte latent collagenase activatable in a phagocytosis-simulated respiratory burst [Tschesche, H. and Macartney, H. W. (1981) Eur. J. Biochem. 120, 183-190].
一种能够抑制多种哺乳动物组织胶原酶的β1 - 血清成分β1 - 抗胶原酶，通过凝胶过滤、亲和色谱和离子交换色谱从人血浆中分离出来。该抑制剂含有1 - 2个游离巯基，这是抑制活性以及与巯基 - 琼脂糖亲和支持物结合的先决条件。碘乙酰胺对β1 - 抗胶原酶的烷基化作用会阻断其抑制活性。该抑制剂被纯化至表观均一性，通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳测定其相对分子质量为30500。测定了其氨基酸和碳水化合物组成。根据其组成和等电聚焦结果，β1 - 抗胶原酶是一种酸性蛋白，等电点为5.6。对人白细胞胶原酶的抑制以强烈的1:1化学计量反应进行。这种结合机制通过二硫键/巯基交换反应发生，正如之前在人白细胞胶原酶形成潜伏酶时所表明的那样[麦卡特尼，H. W. 和切舍，H.（1980年）欧洲生物化学学会联合会快报119，327 - 332]。β1 - 抗胶原酶 - 白细胞胶原酶复合物（潜伏酶）可被含二硫键的化合物如胱氨酸、氧化型谷胱甘肽、胰岛素、松弛素、胰蛋白酶原等激活，但不能被179,203 - 二（S - 羧甲基）胰蛋白酶原或其胰蛋白酶衍生物激活。含有无法接近的二硫键的化合物，如胰凝乳蛋白酶，或巯基的化合物，如D - 青霉胺，不能激活该复合物。然而，如之前在模拟吞噬作用呼吸爆发中可激活的人白细胞潜伏胶原酶所证明的那样，使用产生氧化型谷胱甘肽的系统髓过氧化物酶/H₂O₂/谷胱甘肽很容易实现激活[切舍，H. 和麦卡特尼，H. W.（1981年）欧洲生物化学杂志120，183 - 190]。
