使用C端生物素标记的短杆菌肽A检测膜上的蛋白质-配体相互作用。

Detection of protein-ligand interaction on the membranes using C-terminus biotin-tagged alamethicin.

作者信息

Zhang Y, Futaki Shiroh, Kiwada Tatsuto, Sugiura Yukio

机构信息

Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.

出版信息

Bioorg Med Chem. 2002 Aug;10(8):2635-9. doi: 10.1016/s0968-0896(02)00105-0.

DOI:10.1016/s0968-0896(02)00105-0

PMID:12057652

Abstract

C-terminal biotin-tagged alamethicin, which has several alpha-aminoisobutyric acid (Aib) residues in its sequence, was synthesized by the preparation of the protected peptide segment using the 2-chlorotrityl resin, followed by conjugation with biotin hydrazide. Suppression of the channel current of the biotin-tagged alamethicin by the addition of streptavidin to the electrolyte was monitorable in real time using the planar lipid-bilayer method. The system was also applicable to the detection of interaction of the biotin-tagged alamethicin with the anti-biotin antibody.

摘要

C 端生物素标记的短杆菌肽 A，其序列中有几个α-氨基异丁酸（Aib）残基，通过使用 2-氯三苯甲基树脂制备受保护的肽段，然后与生物素酰肼偶联来合成。使用平面脂质双层方法可实时监测向电解质中添加链霉亲和素后生物素标记的短杆菌肽 A 的通道电流抑制情况。该系统也适用于检测生物素标记的短杆菌肽 A 与抗生物素抗体的相互作用。