三丁基硅异硫氰酸酯作偶联试剂进行蛋白质和多肽C端序列分析

C-terminal Sequence Analysis of Proteins and Peptides Using Tributylsilyl Isothiocyanate

报道了一种新的用于化学降解法蛋白质C 端序列分析的偶联试剂三丁基硅异硫氰酸酯 (tributylsilylisothiocyanate,TBuS ITC) .用氯化三丁基硅和硫氰酸钾反应生成三丁基硅异硫氰酸酯 ,然后以乙酸酐作活化试剂 ,TBuS ITC乙腈溶液作偶联试剂 ,以合成的 10肽为模型肽 ,在低纳摩尔的水平上测到了该肽的C端 4个氨基酸 .其初始产率为 5 3 .1% ,重复产率为 5 7.6% .

A new derivatization reagent, tributylsilyl isothiocyanate(TBuS-ITC), is applied to C-terminal sequence analysis of peptides. The chemistry involves activation with acetic anhydride, derivatization with TBuS-ITC and cleavage of derivatized C-terminal amino acid thiohydantoin with sodium hydroxide. Four C-terminal residues of a synthetic peptide at low nanomole levels have been identified. The initial yields is 53.1% and the repetitive yield is 57.6%.