Chaperonin-encapsulation of proteins for NMR.
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.
Tanaka
Shinji
S
Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Kawata
Yasushi
Y
Otting
Gottfried
G
Dixon
Nicholas E
NE
Matsuzaki
Katsumi
K
Hoshino
Masaru
M
D018835:Chaperonin 10 / Q000737:chemistry; D018834:Chaperonin 60 / Q000737:chemistry; D018833:Chaperonins / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry
D018835:Chaperonin 10; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; D046912:Multiprotein Complexes; D011506:Proteins; D025801:Ubiquitin; D018833:Chaperonins