用4-对二甲氨基偶氮苯-4'-异硫氰酸酯对肽和蛋白质进行高灵敏度序列分析。
High-sensitivity sequence analysis of peptides and proteins by 4-NN-dimethylaminoazobenzene 4'-isothiocyanate.
作者信息
Chang J Y
出版信息
Biochem J. 1977 Jun 1;163(3):517-20. doi: 10.1042/bj1630517.
Abstract
A manual high-sensitivity sequencing method is described, in which 4-NN-dimethylaminoazobenzene 4'-isothiocyanate is used for the stepwise degradation of amino acid residues from the peptides. The 4-NN-dimethylaminoazobenzene 4'-thiazolinones of amino acids that were released, after conversion into their thiohydantoin derivatives, were identified by t.l.c. on polyamide sheets. This new method is simple and sensitive, and requires only 2-10nmol of peptides or proteins for extended sequence analysis. The method was tested on the sequence analysis of a hexapeptide (Leu-Trp-Met-Arg-Phe-Ala), bradykinin, glucagon and native lysozyme. Results show that the proposed procedure is a sensitive method for the sequence determination of short peptides as well as for the partial sequence determination of intact proteins.
摘要
本文描述了一种手动高灵敏度测序方法，其中使用4-N-二甲基氨基偶氮苯4'-异硫氰酸酯对肽段中的氨基酸残基进行逐步降解。释放出的氨基酸的4-N-二甲基氨基偶氮苯4'-噻唑啉酮在转化为硫代乙内酰脲衍生物后，通过在聚酰胺薄板上的薄层层析进行鉴定。这种新方法简单且灵敏，进行扩展序列分析仅需2 - 10 nmol的肽或蛋白质。该方法已用于六肽（亮氨酸-色氨酸-甲硫氨酸-精氨酸-苯丙氨酸-丙氨酸）、缓激肽、胰高血糖素和天然溶菌酶的序列分析。结果表明，所提出的方法对于短肽的序列测定以及完整蛋白质的部分序列测定是一种灵敏的方法。
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