4-NN-二甲基氨基偶氮苯-4'-异硫氰酸盐，一种用于蛋白质序列分析的新型发色试剂。
4-NN-dimethylaminoazobenzene 4'-isothiocyanate, a new chromophoric reagent for protein sequence analysis.
作者信息
Chang J Y, Creaser E H, Bentley K W
出版信息
Biochem J. 1976 Mar 1;153(3):607-11. doi: 10.1042/bj1530607.
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.
合成4-NN-二甲基氨基偶氮苯4'-异硫氰酸酯是为了提高肽序列分析的简便性和灵敏度。描述了4-NN-二甲基氨基偶氮苯4'-异硫氰酸酯的合成方法、24种氨基酸的4-NN-二甲基氨基偶氮苯-4'-硫代乙内酰脲的制备及其薄层层析分离。除亮氨酸和异亮氨酸的硫代乙内酰脲外，所有硫代乙内酰脲都能在二维聚酰胺薄板上通过色谱法得到满意的分离。灵敏的偶氮基团可直接在薄板上检测到皮摩尔量的氨基酸4-NN-二甲基氨基偶氮苯-4'-硫代乙内酰脲呈红色斑点。还报道了一种用于二肽和蛋白质前两个或三个N端氨基酸测序的简单灵敏方法。斑点在暴露于HCl蒸汽后从紫色变为蓝色再变为红色的颜色变化，对应于从4-NN-二甲基氨基偶氮苯-4'-异硫氰酸酯到4-NN-二甲基氨基偶氮苯-4'-硫代氨基甲酰氨基酸衍生物再到4-NN-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，揭示了该试剂一个非常有趣且有价值的特性。
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