Sakac D, Zachos M, Lingwood C A
Research Institute, Hospital for Sick Children, Ontario, Canada.
J Biol Chem. 1992 Jan 25;267(3):1655-9.
The rat testicular galactolipid sulfotransferase has been purified by affinity chromatography using 3'5'-adenosine diphosphate-agarose affinity chromatography. Both galactosyl glycerolipid and galactosyl ceramide were effective substrates with Km values of 4.8 and 1.1 microM respectively. A single protein of molecular mass 56 kDa was present in the purified sulfotransferase preparation as monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining. Specific photoaffinity substrate labeling, using an azido derivative of galactosyl ceramide, was used to identify this protein, both in crude extracts and when purified. The protein was also selectively phosphorylated in the presence of the rat testicular galactolipid sulfotransferase stimulatory protein kinase.
大鼠睾丸半乳糖脂磺基转移酶已通过使用3'5'-二磷酸腺苷-琼脂糖亲和色谱的亲和色谱法进行了纯化。半乳糖基甘油脂和半乳糖基神经酰胺都是有效的底物，其Km值分别为4.8和1.1微摩尔。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和银染监测，纯化的磺基转移酶制剂中存在一种分子量为56 kDa的单一蛋白质。使用半乳糖基神经酰胺的叠氮衍生物进行特异性光亲和底物标记，用于在粗提物和纯化时鉴定该蛋白质。在大鼠睾丸半乳糖脂磺基转移酶刺激蛋白激酶存在的情况下，该蛋白质也被选择性磷酸化。
