Prehm P, Mausolf A
Biochem J. 1986 May 1;235(3):887-9. doi: 10.1042/bj2350887.
Hyaluronate synthase was isolated from protoblast membranes of streptococci by Triton X-114 extraction and cetylpyridinium chloride precipitation. It was identified as a 52,000-Mr protein, which bound to nascent hyaluronate and was affinity-labelled by periodate-oxidized UDP-glucuronic acid and UDP-N-acetylglucosamine. Antibodies directed against the 52,000-Mr protein inhibited hyaluronate synthesis. Mutants defective in hyaluronate synthase activity lacked the 52,000-Mr protein in membrane extracts. Synthase activity was solubilized from membranes by cholate in active form and purified by ion-exchange chromatography.
通过Triton X-114提取和十六烷基氯化吡啶沉淀从链球菌的原生质膜中分离出透明质酸合酶。它被鉴定为一种分子量为52,000的蛋白质，该蛋白质与新生的透明质酸结合，并被高碘酸盐氧化的UDP-葡萄糖醛酸和UDP-N-乙酰葡糖胺进行亲和标记。针对分子量为52,000的蛋白质的抗体抑制透明质酸的合成。透明质酸合酶活性有缺陷的突变体在膜提取物中缺乏分子量为52,000的蛋白质。合酶活性通过胆酸盐以活性形式从膜中溶解出来，并通过离子交换色谱法进行纯化。
