Chaperonin-encapsulation of proteins for NMR

Authors

Shinji Tanaka, Kyoto University
Yasushi Kawata, Tottori University
Gottfried Otting, Australian National University
Nicholas E. Dixon, University of WollongongFollow
Katsumi Matsuzaki, Kyoto University
Masaru Hoshino, Kyoto University

RIS ID

32931

Publication Details

Matsuzaki, K., Kawata, Y., Hoshino, M., Dixon, N. E., Otting, G. & Tanaka, S. (2010). Chaperonin-encapsulation of proteins for NMR. BBA - Proteins and Proteomics, 1804 (4), 866-871.

Abstract

A novel chaperonin-encapsulation system for NMR measurements has been designed. The single-ring variant SR398 with an ATPase deficient mutation of GroEL, also known as chaperonin, bound co-chaperonin GroES irreversibly, forming a stable cage to encapsulate a target protein. A small GroEL-binding tag made it possible to perform all steps of the encapsulation under near physiological conditions while retaining the native conformation of the target protein. About half of the SR398/GroES cages encapsulated target protein molecules. As binding only depends on the 12-residue tag sequence, this encapsulation method is applicable to a large number of proteins. Isolation of the target proteins in the molecular cage of chaperonin will allow the study of highly aggregation-prone proteins by solution NMR.