Egorov T A, Svenson A, Rydén L, Carlsson J
Proc Natl Acad Sci U S A. 1975 Aug;72(8):3029-33. doi: 10.1073/pnas.72.8.3029.
Activated thiol-Sepharose [agarose-(glutathione-2-pyridyl disulfide) conjugate] has been used to immobilize proteins with a single or a few thiol groups via disulfide bridges. The immobilized proteins were subsequently proteolytically degraded. After washing, the thiol-containing peptides were eluted with a reducing agent. A single preparative paper electrophoresis, occasionally after a modification such as oxidation, was sufficient to obtain pure peptides in good yields. The method was applied to the major parvalbumin from hake muscle (a protein with 108 amino acid residues and one cysteine residue), to mercaptalbumin from bovine serum (565 residues and one cysteine), and to human serum ferroxidase [EC 1.16.3.1; iron (II):oxygen oxidoreductase] (ceruloplasmin) (1065 residues and three cysteines). The use of the technique, e.g., as a simple means of obtaining homologous peptides in related proteins, is discussed.
活化硫醇琼脂糖凝胶[琼脂糖-（谷胱甘肽-2-吡啶二硫化物）偶联物]已被用于通过二硫键固定含有单个或几个硫醇基团的蛋白质。随后对固定化的蛋白质进行蛋白水解降解。洗涤后，用还原剂洗脱含硫醇的肽段。单次制备型纸电泳，偶尔在经过如氧化等修饰后，就足以以良好的产率获得纯肽段。该方法应用于无须鳕肌肉中的主要副肌球蛋白（一种含有108个氨基酸残基和一个半胱氨酸残基的蛋白质）、牛血清中的巯基白蛋白（565个残基和一个半胱氨酸）以及人血清铁氧化酶[EC 1.16.3.1；铁（II）：氧氧化还原酶]（铜蓝蛋白）（1065个残基和三个半胱氨酸）。本文还讨论了该技术作为一种获取相关蛋白质中同源肽段的简单方法的应用。
