Photoaffinity labeling of insulin receptor proteins of liver plasma membrane preparations.
作者： Cecil C. Yip , Clement W. T. Yeung , Margaret L. Moule
DOI: 10.1021/BI00542A011
关键词: Insulin receptor 、 Photoaffinity labeling 、 Molecular mass 、 Insulin receptor substrate 、 Gel electrophoresis 、 Insulin 、 Biochemistry 、 Chemistry 、 Sodium dodecyl sulfate 、 Polyacrylamide gel electrophoresis
摘要: The photoreactive insulin derivatives N epsilon B29-(azidobenzoyl)insulin (MAB-insulin) and alpha A1, B29-di(azidobenzoyl)insulin (DAB-insulin) were synthesized by reacting bovine with the N-hydroxysuccinimide ester of 4-azidobenzoic acid. These purified ion exchange chromatography on SP-Sephadex, their identities established polyacrylamide gel electrophoresis, amino acid analysis, end-group determination. Their biological activities measured receptor binding assay fat cell assay. photoreactivity these two was demonstrated spectral changes formation covalent polymers high molecular weight when exposed to light. Radioactive MAB-insulin DAB-insulin prepared iodination [125I]iodine. radioactive characterized for photoreactivity, immunoreactivity, liver plasma membrane. Liver membrane preparations rat, mouse, guinea pig incubated irradiated Sodium dodecyl sulfate electrophoresis after solubilization reduction showed that proteins specifically labeled. weights bands estimated be about 130 000 90 in all three species animals.
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