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:1972
卷号:69
期号:5
页码:1277-1281
DOI:10.1073/pnas.69.5.1277
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:Relatively simple and rapid procedures are described for the large-scale preparation of liver membranes that contain virtually all of the high affinity insulin-binding activity of liver homogenates. The presumed insulin recepotr, which is extracted from these membranes in soluble form with Triton X-100, can be further purified by ammonium sulfate fractionation (3-fold purification) or by diethylaminoethyl-cellulose chromatography (60-fold purification). Several insulin-agarose derivatives have been synthesized that can efficiently extract the insulin-binding protein from the detergent extracts of the membranes. The receptor macro-molecule can be eluted from the affinity columns in high (50-80%) yield by use of urea-containing buffers of moderately low pH. The receptor, thus purified by small-scale affinity chromatography experiments, approaches theoretical purity on the basis of its specific activity. This protein is purified about 250,000-fold from the liver homogenate by detergent extraction and affinity chromatography.