期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1978
卷号：75
期号：4
页码：1768-1772
DOI：10.1073/pnas.75.4.1768
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Protein synthesized in ascites cell-free extracts in response to first trimester placental mRNA was immunoprecipitated with antisera directed against the subunit of human chorionic gonadotropin. The immunoprecipitable proteins were resolved on sodium dodecyl sulfate/polyacrylamide slab gels. In membrane-depleted extracts placental mRNA directed the synthesis of the "preprotein" form of the subunit. However, when membranes were added to the cell-free extracts a protein migrating more slowly than pre- subunit was observed. This protein was specifically adsorbed to a concanavalin A column, and its migration on sodium dodecyl sulfate gels was enhanced after treatment with -mannosidase (EC 3.2.1.24 ) or endo-{beta}-N-acetylglucosaminidase CII or H. Similar results were obtained when mRNA was translated in lysates derived from first trimester placenta instead of in ascites cell extracts. Kinetic studies of the glycosylation reaction revealed that sugar attachment can occur just prior to release of the protein. These data show that the apoprotein of the subunit can be glycosylated in vitro. The data also suggest that the glycosylated protein contains a sugar core consisting of di-N-acetylchitobiose and at least four mannose residues, some of which are -linked. In addition, it appears that this carbohydrate unit is present in homologous placental membranes as well as in the ascites tumor membranes.
关键词：glycoprotein hormone ; secretory protein ; in vitro protein synthesis ; membrane-dependent sugar transfer
