Liu Y, Liang S
College of Life Sciences, Hunan Normal University, Changsha, People's Republic of China.
J Protein Chem. 2001 Oct;20(7):535-41. doi: 10.1023/a:1013367728843.
A new derivatization reagent, tribenzylsilyl isothiocyanate (TBS-ITC), is applied to C-terminal peptide and protein sequencing. It has been successfully used to sequence six C-terminal residues of house apomyoglobin and a synthetic peptide at low nanomole levels. The chemistry involves activation with acetic anhydride, derivatization with TBS-ITC, and cleavage of derivatized C-terminal amino acid thiohydantoin with sodium hydroxide. The tribenzylsilyl is a bulky, electric donor group and is a good leaving group. It facilitates the nucleophilic attack of the NCS(-1) in the coupling reaction. The efficiency for C-terminal sequencing by TBS-ITC is about the same as that of acetyl isothiocyanate (AITC), which is a derivatizing reagent for C-terminal sequencing developed by our laboratory. TBS-ITC is much more stable than AITC and trimethylsilyl isothiocyanate (TMS-ITC). TBS-ITC is a solid with relatively long shelf life, whereas AITC and TMS-ITC are liquid and not stable at room temperature.
一种新型衍生试剂，三苄基甲硅烷基异硫氰酸酯（TBS-ITC），被应用于C端肽和蛋白质测序。它已成功用于在低纳摩尔水平下对马心脱辅基肌红蛋白的六个C端残基和一个合成肽进行测序。该化学过程包括用乙酸酐活化、用TBS-ITC衍生化，以及用氢氧化钠裂解衍生化的C端氨基酸硫代乙内酰脲。三苄基甲硅烷基是一个庞大的供电子基团，并且是一个良好的离去基团。它在偶联反应中促进NCS(-1)的亲核进攻。用TBS-ITC进行C端测序的效率与乙酰异硫氰酸酯（AITC）大致相同，AITC是我们实验室开发的一种用于C端测序的衍生试剂。TBS-ITC比AITC和三甲基甲硅烷基异硫氰酸酯（TMS-ITC）稳定得多。TBS-ITC是一种具有相对较长保质期的固体，而AITC和TMS-ITC是液体且在室温下不稳定。
