SEPARATION AND CHARACTERIZATION OF CHONDROITIN 6-SULFOTRANSFERASE AND CHONDROITIN 4-SULFOTRANSFERASE FROM CHICK-EMBRYO CARTILAGE
SEPARATION AND CHARACTERIZATION OF CHONDROITIN 6-SULFOTRANSFERASE AND CHONDROITIN 4-SULFOTRANSFERASE FROM CHICK-EMBRYO CARTILAGE
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DOI：
10.1016/0304-4165(82)90282-3
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发表时间：
1982-01-01
期刊：
影响因子：
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通讯作者：
MIYASHITA, N
中科院分区：
文献类型：
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作者：
HABUCHI, O;MIYASHITA, N
从 14 日龄鸡胚的骨骺软骨中提取两种不同的磺基转移酶（软骨素 6-磺基转移酶和软骨素 4-磺基转移酶），它们催化硫酸盐转移到软骨素乙酰半乳糖胺残基的位置 6 和位置 4，并在 3 M 存在的情况下通过 Sephacryl S-200 凝胶色谱进行分离。盐酸胍。当含有 3 M 盐酸胍的酶溶液在含有 10% 甘油的 0.02 M Tris-HCl（pH 7.2）中透析时，软骨素 4-磺基转移酶变得几乎不溶；软骨素6-磺基转移酶保持可溶。从两种酶制剂中完全去除了硫酸盐转移的内源受体。向培养介质中添加碱性蛋白质和多胺以及 Mn2+ 会刺激两种磺基转移酶；这些阳离子对软骨素6-磺基转移酶的刺激高于软骨素4-磺基转移酶。两种酶的 3''-磷酸腺苷基硫酸盐的 Km 值在鱼精蛋白或精胺存在下比在 Mn2+ 存在下小得多。两种磺基转移酶对巯基化合物的需求不同；在没有巯基化合物的情况下，软骨素4-磺基转移酶的活性很低；软骨素6-磺基转移酶的活性基本不受影响。至少有 2 种磺基转移酶参与硫酸软骨素的生物合成。软骨细胞中硫酸软骨素异构体的产生受到多种因素的影响，例如细胞内的巯基化合物和碱性物质的浓度。
Two distinct sulfotransferases (chondroitin 6-sulfotransferase and chondroitin 4-sulfotransferase), which catalyzed transfer of sulfate to position 6 and position 4 ofacetylgalactosamine residues of chondroitin, were extracted from epiphyseal cartilage of 14-day-old chick embryos and separated by gel chromatography on Sephacryl S-200 in the presence of 3 M guanidine-HCl. When the enzyme solutions containing 3 M guanidine-HCl were dialyzed against 0.02 M Tris-HCl, pH 7.2, containing 10% glycerol, chondroitin 4-sulfotransferase became almost insoluble; chondroitin 6-sulfotransferase remained soluble. Endogenous acceptors for sulfate transfer were completely removed from both enzyme preparations. Addition of basic proteins and polyamines as well as Mn2+ to the incubation medium caused a stimulation of both sulfotransferases; the stimulation of chondroitin 6-sulfotransferase with these cations was higher than that of chondroitin 4-sulfotransferase. Km values for 3''-phosphoadenylyl sulfate of both enzymes were much smaller in the presence of protamine or spermine than in the presence of Mn2+. The 2 sulfotransferases differed in the requirement for sulfhydryl compounds; in the absence of sulfhydryl compounds, the activity of chondroitin 4-sulfotransferase was very low; the activity of chondroitin 6-sulfotransferase was essentially unaffected. At least 2 sulfotransferases are involved in the biosynthesis of chondroitin sulfate. Production of the isomers of chondroitin sulfate in chondrocytes is affected by various factors such as the intracellular concentration of sulfhydryl compounds and basic substances.