Li M, Distler J J, Jourdian G W
Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor 48109.
Glycobiology. 1991 Nov;1(5):511-7. doi: 10.1093/glycob/1.5.511.
A convenient means was devised for the purification of milligram quantities of a soluble form of the mannose 6-phosphate/insulin-like growth factor II receptor (Man-6-P/IGF II receptor). The receptor was purified to near homogeneity from bovine serum by affinity chromatography on agarose-pentamannosephosphate in the absence of detergent. Approximately 2.5 mg of receptor were obtained from 500 ml of fetal calf serum. The concentration of receptor in serum decreased sharply with development. Fetal calf serum Man-6-P/IGF II receptor was immunologically similar to detergent-solubilized, membrane-bound Man-6-P/IGF II receptor from bovine liver. N-Terminal sequence analysis revealed that the purified serum receptor, but not the solubilized, membrane-associated receptor, contains stoichiometric amounts of bound IGF II. The results of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel chromatography studies suggest that the fetal calf serum receptor (in contrast to the solubilized, membrane-bound bovine testis receptor) does not aggregate. The affinity of the fetal calf serum receptor for bovine testis beta-galactosidase approximated one-half that observed for solubilized, membrane-bound bovine testis receptor.
已设计出一种简便方法，用于纯化毫克量的可溶性甘露糖6-磷酸/胰岛素样生长因子II受体（Man-6-P/IGF II受体）。在无去污剂的情况下，通过琼脂糖-五甘露糖磷酸亲和色谱法从牛血清中纯化该受体，使其接近均一。从500毫升胎牛血清中大约获得2.5毫克受体。血清中受体的浓度随着发育而急剧下降。胎牛血清中的Man-6-P/IGF II受体在免疫上与来自牛肝的经去污剂溶解的膜结合Man-6-P/IGF II受体相似。N端序列分析表明，纯化的血清受体而非溶解的膜相关受体含有化学计量的结合IGF II。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和凝胶色谱研究结果表明，胎牛血清受体（与溶解的膜结合牛睾丸受体相反）不会聚集。胎牛血清受体对牛睾丸β-半乳糖苷酶的亲和力约为溶解的膜结合牛睾丸受体的一半。
