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Attachment of an NMR-invisible solubility enhancement tag using a sortase-mediated protein ligation method

  • Yoshihiro Kobashigawa
  • Hiroyuki Kumeta
  • Kenji Ogura
  • Fuyuhiko Inagaki
Communication

Abstract

Sample solubility is essential for structural studies of proteins by solution NMR. Attachment of a solubility enhancement tag, such as GB1, MBP and thioredoxin, to a target protein has been used for this purpose. However, signal overlap of the tag with the target protein often made the spectral analysis difficult. Here we report a sortase-mediated protein ligation method to eliminate NMR signals arising from the tag by preparing the isotopically labeled target protein attached with the non-labeled GB1 tag at the C-terminus.

Keywords

Sortase Protein ligation GB1 Intein INSET Solubility enhancement 

Abbreviations

SDS-PAGE

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

INSET

Isotopically invisible solubility/stability enhancement tag

Notes

Acknowledgements

This work was supported by Grants-in-Aid for Scientific Research and the National Project on “Targeted Proteins Research Program” from the Ministry of Education, Science and Culture of Japan.

Supplementary material

10858_2008_9296_MOESM1_ESM.pdf (433 kb)
Supplementary material 1 (PDF 432 kb)

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Yoshihiro Kobashigawa
    • 1
  • Hiroyuki Kumeta
    • 1
  • Kenji Ogura
    • 1
  • Fuyuhiko Inagaki
    • 1
  1. 1.Department of Structural Biology, Graduate School of Pharmaceutical SciencesHokkaido UniversitySapporoJapan

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