Ferl R J
Biochem J. 1985 Mar 15;226(3):853-8. doi: 10.1042/bj2260853.
Experiments utilizing proteolytic mapping of maize Alcohol dehydrogenase-1 protein (EC 1.1.1.1; ADH) showed that, in the presence of sodium dodecyl sulphate (SDS), two discrete areas of the protein molecule were hypersensitive to digestion with Staphylococcus aureus V8 proteinase. These areas were mapped to points 11 and 27 kDa along the 41 kDa polypeptide. Furthermore, ADH1 proteins isolated from the ethyl methanesulphonate-induced mutants U725 and W586 showed both a change in electrophoretic mobility in SDS gels, and an altered V8 proteinase digestion pattern. Protein from U725 migrated in SDS gels as though it was 2 kDa smaller than wild-type ADH protein and lacked the 11 kDa cleavage site. Protein from W586 lacked the 30 kDa cleavage site and migrated as if it was 3 kDa smaller than wild type. The possible relationships between proteinase cleavage sites, mutations and SDS gel mobilities are discussed.
利用玉米乙醇脱氢酶-1蛋白（EC 1.1.1.1；ADH）的蛋白水解图谱进行的实验表明，在十二烷基硫酸钠（SDS）存在的情况下，该蛋白分子的两个离散区域对金黄色葡萄球菌V8蛋白酶的消化高度敏感。这些区域在41 kDa多肽上分别定位到11 kDa和27 kDa处。此外，从甲磺酸乙酯诱导的突变体U725和W586中分离出的ADH1蛋白在SDS凝胶中的电泳迁移率发生了变化，并且V8蛋白酶的消化模式也有所改变。U725的蛋白在SDS凝胶中的迁移情况表明它比野生型ADH蛋白小2 kDa，并且缺少11 kDa的切割位点。W586的蛋白缺少30 kDa的切割位点，其迁移情况表明它比野生型小3 kDa。本文讨论了蛋白酶切割位点、突变与SDS凝胶迁移率之间可能的关系。
