Staswick P E, Hermodson M A, Nielsen N C
J Biol Chem. 1984 Nov 10;259(21):13431-5.
The half-cystine residues involved in linking the acidic and basic polypeptides were determined for several glycinin subunits. The cystines were localized with specific cyanogen bromide fragments either by comparing the electrophoretic mobility of nonreduced and reduced fragments, or by co-purifying and then determining the NH2-terminal sequence of the covalently linked fragments. Residues involved in the disulfides were further identified by labeling them with [3H] iodoacetic acid. Only 1 cystine was found to be involved in linking the acidic and basic components of each subunit, and they were in analogous positions in each of the subunits studied. Potential sites for intrapolypeptide cystines were also identified.
已针对几种大豆球蛋白亚基确定了参与连接酸性和碱性多肽的半胱氨酸残基。通过比较非还原片段和还原片段的电泳迁移率,或通过共纯化然后确定共价连接片段的NH2末端序列,用特定的溴化氰片段定位胱氨酸。通过用[3H]碘乙酸标记参与二硫键的残基,进一步对其进行了鉴定。发现每个亚基的酸性和碱性成分之间仅由1个胱氨酸连接,并且在所研究的每个亚基中它们处于类似位置。还确定了多肽内胱氨酸的潜在位点。