Chemical & Pharmaceutical Bulletin 2014-01-01

Specific labeling of streptavidin for better understanding of ligand modification in modular method for affinity labeling (MoAL).

Munetaka Kunishima, Daiki Kato, Shuichi Nakanishi, Masanori Kitamura, Kohei Yamada, Keiji Terao, Tomoya Asano

文献索引：Chem. Pharm. Bull. 62(11) , 1146-50, (2014)

全文：HTML全文

摘要

We studied the specific labeling of streptavidin using the modular method for affinity labeling (MoAL) that we developed based on a catalytic amide-forming reaction using 2-chloro-4,6-dimethoxy-1,3,5-triazine (CDMT) and a tertiary amine catalyst. The primary structures of avidin and streptavidin are significantly different from each other, and streptavidin does not possess an acidic amino acid equivalent to Asp108 of avidin, which is the target acidic amino acid that was labeled using MoAL. However, using biotinylated modular ligand catalysts (MLC) originally designed for labeling avidin, the labeling of streptavidin was found to successfully proceed at Glu51, which is located in a different region. The present study indicates that MoAL is readily applicable to protein labeling without a precise design for MLC. The most important factor for the design of MLC is to ensure that the linker is of sufficient length to connect the ligands to a catalytic site.