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标题：Cell-free translation and partial characterization of mRNA coding for enkephalin-precursor protein
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作者：S Dandekar ; S L Sabol
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：1982
卷号：79
期号：4
页码：1017-1021
DOI：10.1073/pnas.79.4.1017
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：The biosynthesis of the protein precursor of [Met]enkephalin (Tyr-Gly-Gly-Phe-Met) was studied using cell-free translation systems to characterize the enkephalin-precursor gene product and mRNA. Proteins obtained by translation of bovine adrenal medullary mRNA in the presence of [35S]methionine were digested with trypsin and carboxypeptidase B. The resultant peptides were immunoprecipitated with anti[Met]enkephalin serum and subsequently analyzed by reverse phase HPLC. [35S][Met]Enkephalin (up to 0.2 fmol/micrograms mRNA, identified by antigenic specificity and chromatographic mobility, was isolated from peptides obtained from proteins whose synthesis was dependent upon adrenal medullary mRNA. Both trypsin and carboxy-peptidase B were required to generate [35S][Met]enkephalin from translation products. The largest protein containing the [35S]-[Met]enkephalin sequence synthesized in the wheat germ system has a Mr, of 31,000 +/- 1000, determined by NaDodSO4/polyacrylamide gel electrophoresis. Adrenal medullary mRNA coding for protein containing the [35S][Met]enkephalin sequence was resolved into a major fraction of 1450 +/- 150 nucleotides and a minor fraction of 47000 +/- 450 nucleotides, as determined by agarose gel electrophoresis in the presence of methylmercuric hydroxide. It is proposed that the major enkephalin-precursor gene product of adrenal medulla is a protein of Mr approximately 31,000.U