title: Chaperonin-encapsulation of proteins for NMR.
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.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Tanaka
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kawata
authors.forename: Gottfried
authors.initials: G
authors.lastname: Otting
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Dixon
authors.forename: Katsumi
authors.initials: K
authors.lastname: Matsuzaki
authors.forename: Masaru
authors.initials: M
authors.lastname: Hoshino
mesh_terms: 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
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; D046912:Multiprotein Complexes; D011506:Proteins; D025801:Ubiquitin; D018833:Chaperonins