Utarabhand P, Bunlipatanon P
Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Thailand.
Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1996 Oct;115(2):101-10. doi: 10.1016/s0742-8413(96)00055-2.
Radiolabelled vitellogenin produced by juvenile grouper following injection of 3H-leucine, 32P-orthophosphate and estradiol-stimulation was purified by anion-exchange and gel filtration chromatography and characterized by electrophoresis and chemical analysis. The radiolabelled product was found to exist in two heterogeneous molecular weight forms by electrophoresis under nondenaturing conditions on polyacrylamide gel with M(r), of 260,000 and 525,000. It showed two protein monomers (M(r) 113,000 and 140,000) on electrophoresis under denaturing condition on a sodium dodecyl sulphate-polyacrylamide gel. The purified material contained 17.2% total lipid with 1.3% cholesterol and 2.1% triglycerides, 8-21 micrograms/mg protein total carbohydrate and 6.8 micrograms phosphate/mg protein; amino acid analysis showed a profile comparable to that found for vitellogenins isolated from the grouper, medaka, goldfish and rainbow trout. The study showed grouper vitellogenin to be a glycophospholipoprotein similar in composition to vitellogenins from other teleosts and demonstrated that vitellogenins can be induced in juveniles by injection with estradiol-17 beta.
通过注射³H-亮氨酸、³²P-正磷酸盐并经雌二醇刺激后，由幼体石斑鱼产生的放射性标记卵黄蛋白原，经阴离子交换和凝胶过滤色谱法纯化，并通过电泳和化学分析进行表征。在非变性条件下于聚丙烯酰胺凝胶上进行电泳时，发现该放射性标记产物以两种不同分子量形式存在，分子量分别为260,000和525,000。在十二烷基硫酸钠-聚丙烯酰胺凝胶上进行变性条件下的电泳时，它显示出两种蛋白质单体（分子量分别为113,000和140,000）。纯化后的物质含有17.2%的总脂质，其中胆固醇占1.3%，甘油三酯占2.1%，每毫克蛋白质含8 - 21微克总碳水化合物，每毫克蛋白质含6.8微克磷酸盐；氨基酸分析显示其图谱与从石斑鱼、青鳉、金鱼和虹鳟中分离出的卵黄蛋白原的图谱相当。该研究表明，石斑鱼卵黄蛋白原是一种糖磷脂蛋白，其组成与其他硬骨鱼的卵黄蛋白原相似，并证明通过注射17β-雌二醇可在幼体中诱导产生卵黄蛋白原。
