HIGH CONFIDENCE PROTEIN IDENTIFICATION OF PROTEIN ISOLATED BY SDS-PAGE

通过MALDI的肽图谱已长期用于蛋白质 通过SDS-PAGE鉴定分离蛋白。 的有效 肽作图和随后提取含有 半胱氨酸我们研究了几种烷基化剂的使用。 此外，我们已经表明，使用标记剂或 同时使用两种烷基化试剂可以鉴定 在所有胰蛋白酶肽中含有半胱氨酸的肽。 这 信息可用于约束数据库中的搜索， 蛋白质未知。 该方法已得到应用，并显示有效 选择蛋白质。 我们现在正在应用这一方法， 鉴定从核中纯化的蛋白质的方法 信封.

Peptide mapping by MALDI has been long used for protein identification of protein isolated by SDS-PAGE. For an efficient peptide mapping and consequent extraction of peptides containing cysteine we have investigated the use of several alkylating agents. Furthermore, we have shown that the use of labeled agents or the simultaneous use of two alkylating reagents allowed the identification of peptides containing cysteines among all the tryptic peptides. This information can be used to constrain the search in the data base if the protein is unknown. The methodology was applied and show to work with selected proteins. We are now in the process of applying this methods for the identification of proteins purified from the nuclear envelope.