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Chaperonin-encapsulation of proteins for NMR

Journal Article


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. (C) 2010 Elsevier B.V. All rights reserved.

Authors


  •   Tanaka, Shinji (external author)
  •   Kawata, Yasushi (external author)
  •   Otting, Gottfried (external author)
  •   Dixon, Nicholas E.
  •   Matsuzaki, Katsumi (external author)
  •   Hoshino, Masaru (external author)

Publication Date


  • 2010

Citation


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

Scopus Eid


  • 2-s2.0-76849111130

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/5056

Number Of Pages


  • 5

Start Page


  • 866

End Page


  • 871

Volume


  • 1804

Issue


  • 4

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. (C) 2010 Elsevier B.V. All rights reserved.

Authors


  •   Tanaka, Shinji (external author)
  •   Kawata, Yasushi (external author)
  •   Otting, Gottfried (external author)
  •   Dixon, Nicholas E.
  •   Matsuzaki, Katsumi (external author)
  •   Hoshino, Masaru (external author)

Publication Date


  • 2010

Citation


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

Scopus Eid


  • 2-s2.0-76849111130

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/5056

Number Of Pages


  • 5

Start Page


  • 866

End Page


  • 871

Volume


  • 1804

Issue


  • 4