A DIAGONAL PROCEDURE FOR ISOLATING SULFHYDRYL PEPTIDES ALKYLATED WITH N-ETHYLMALEIMIDE
A DIAGONAL PROCEDURE FOR ISOLATING SULFHYDRYL PEPTIDES ALKYLATED WITH N-ETHYLMALEIMIDE
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DOI：
10.1016/0003-2697(80)90396-6
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发表时间：
1980-01-01
影响因子：
2.9
通讯作者：
CHRISTEN, P
中科院分区：
文献类型：
--
作者：
GEHRING, H;CHRISTEN, P
N-乙基马来酰亚胺的烷基化产物虽然在酸性pH下稳定，但在碱性pH下水解成相应的N-乙基琥珀酰胺酸。基于这种行为，开发了用于从蛋白水解消化物的大量未标记肽中分离用N-乙基马来酰亚胺烷基化的巯基肽的对角电泳程序。在pH 6.5下进行第一次高压纸电泳后，将纸条暴露于氨气氛中。N-乙基琥珀酰亚胺部分水解为N-乙基琥珀酰胺酸产生新的负电荷，允许在第二次电泳运行中将标记的肽与未标记的肽容易分离。
The alkylation product of N-ethylmaleimide, while stable at acid pH, is hydrolyzed at alkaline pH to the corresponding N-ethylsuccinamic acid. Based on this behavior, a diagonal electrophoretic procedure for separating sulfhydryl peptides alkylated with N-ethylmaleimide from the bulk of unlabeled peptides of a proteolytic digest was developed. After a 1st high-voltage paper electrophoresis at pH 6.5 the paper strip was exposed to an ammonia atmosphere. Hydrolysis of the N-ethylsuccinimide moiety to N-ethylsuccinamic acid generated a new negative charge allowing ready separation of the labeled from the unlabeled peptides in a 2nd electrophoretic run.