4-NN-dimethylaminoazobenzene 4'-isothiocyanate, a new chromophoric reagent for protein sequence analysis.
4-NN-dimethylaminoazobenzene 4'-isothiocyanate, a new chromophoric reagent for protein sequence analysis.
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4-NN-二甲基氨基偶氮苯 4-异硫氰酸酯，一种用于蛋白质序列分析的新型发色试剂。
DOI：
10.1042/bj1530607
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发表时间：
1976
期刊：
影响因子：
--
通讯作者：
K. W. Bentley
中科院分区：
文献类型：
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作者：
J. Chang;E. H. Creaser;K. W. Bentley
合成了4-N-二甲氨基偶氮苯4 '-异硫氰酸酯，以提高肽序列分析的简便性和灵敏度。本文介绍了4-N-二甲氨基偶氮苯-4 '-异硫氰酸酯的合成方法，24种氨基酸的4-N-二甲氨基偶氮苯-4'-硫代海因的制备及其t.l.c.分离被描述。除亮氨酸和异亮氨酸外，所有的硫代乙内酰脲都能在二维聚酰胺膜上得到满意的分离。灵敏的偶氮基团允许直接在片材上检测4-N，N-二甲基氨基偶氮苯-4 '-硫代乙内酰脲氨基酸的红色斑点，其量低至pmol。还报道了一种简单、灵敏的测定二肽和蛋白质N端前两个或三个氨基酸序列的方法。在暴露于HCl蒸汽后，斑点的颜色从紫色变为蓝色再变为红色，对应于从4-N，N-二甲基氨基偶氮苯-4 '异硫氰酸酯变为4-N，N-二甲基氨基偶氮苯-4'-硫代氨基甲酰基氨基酸衍生物再变为4-N，N-二甲基氨基偶氮苯-4 '-硫代乙内酰脲氨基酸衍生物的化学变化，揭示了该试剂非常有趣和有价值的特征。
4-NN-Dimethylaminoazobenzene 4'-isothiocyanate was synthesized for the purpose of improving the ease and sensitivity of peptide sequence analysis. The method of 4-NN-dimethylaminoazobenzene 4'-isothiocyanate synthesis, the preparation of 24 4-NN-dimethylaminoazobenzene-4'-thiohydantoins of amino acids and their t.l.c. separation are described. All the thiohydantoins, except those of leucine and isoleucine, could be satisfactorily separated by chromatography on a two-dimensional polyamide sheet. The sensitive azo group permits the detection of 4-NN-dimethylaminoazobenzene-4'-thiohydantoins of amino acids as red spots down to pmol amounts directly on the sheet. A simple sensitive method for sequencing dipeptides and the first two or three N-terminal amino acids of proteins is also reported. The colour change of the spots from purple to blue to red after being exposed to HCl vapour, corresponding to the chemical change from 4-NN-dimethylaminoazobenzene-4' isothiocyanate to the 4-NN-dimethylaminoazobenzene-4'-thiocarbamoyl amino acid derivative to the 4-NN-dimethylaminoazobenzene-4'-thiohydantoin amino acid derivative, reveals a very interesting and valuable feature of this reagent.