用硫氰酸盐法从羧基末端对肽段进行测序的研究。
Studies on sequencing of peptides from the carboxyl terminus by using the thiocyanate method.
作者信息
Rangarajan M, Darbre A
出版信息
Biochem J. 1976 Aug 1;157(2):307-16. doi: 10.1042/bj1570307.
Abstract
We report on our experience in applying the thiocyanate method developed by Stark (1968) (Biochemistry 7, 1796-1807) to the sequencing of short peptides from the carboxyl end in free solution. Yields fell to very low levels after three cycles of degradation. The method was time-consuming because of the filtration and freeze-drying stages involved. To overcome these problems, peptides were attached to modified polystyrene polymers for sequential degradation in the solid phase, and a maximum of six amino acids was determined. Also, ribonuclease was attached to active-ester glass beads and sequential degradation was carried out to determine six amino acids at the C-terminal end of this protein.
摘要
我们报告了将斯塔克（1968年）开发的硫氰酸盐法（《生物化学》7，1796 - 1807）应用于在游离溶液中从羧基末端对短肽进行测序的经验。降解三个循环后产率降至非常低的水平。由于涉及过滤和冷冻干燥阶段，该方法很耗时。为克服这些问题，将肽连接到修饰的聚苯乙烯聚合物上以进行固相顺序降解，并确定了最多六个氨基酸。此外，将核糖核酸酶连接到活性酯玻璃珠上并进行顺序降解以确定该蛋白质C末端的六个氨基酸。
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