文章基本信息

标题：Novel Separation and Amino Acid Sequences of α and β Subunits of Pig Heart Pyruvate Dehydrogenase
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作者：Yoshishige URATA ; Kichiko KOIKE ; Shinji GOTO 等
期刊名称：Journal of Nutritional Science and Vitaminology
印刷版ISSN：0301-4800
电子版ISSN：1881-7742
出版年度：1991
卷号：37
期号：3
页码：257-267
DOI：10.3177/jnsv.37.257
出版社：Center for Academic Publications Japan
摘要：Pyruvate dehydrogenase was separated from pig heart pyruvate dehydrogenase complex by gel-permeation chromatography on a TSK G4000SWG column in the presence of 4M urea, followed by chromatography on a calcium phosphate gel-cellulose column. The pyruvate dehydrogenase was further separated into two nonidentical subunits, α and β, by high-performance liquid chromatography on a Synchropak CM-300 column in the presence of 8 M urea. The complete amino acid sequences of two subunits of pyruvate dehydrogenase were determined. The peptide fragments of S -carboxymethylated subunits were generated by treatment with endoproteinase Lys-C, endoproteinase Asp-N, trypsin, and cyanogen bromide. The subunits α and β contain 361 amino acid residues ( M _r 40, 294) and 329 residues ( M _r 35, 787), respectively. The amino acid sequences of subunits α and β in the pig were, respectively, 98 and 96% identical to those in humans. Hydropathy analysis and prediction of the secondary structure of two subunits suggest that the subunit α contains the thiamin pyrophosphate-binding domain and that the subunit β contains segments with a high hydrophobicity.
关键词：amino acid sequence;pyruvate dehydrogenase;pyruvate dehydrogenase complex;TSK G4000SWG column;high-performance liquid chromatography