Ribonuclease H: Full Assignment of Backbone Proton Resonances with Heteronuclear 3D NMR and Solution Structure

  • Kuniaki Nagayama
  • Toshio Yamazaki
  • Mayumi Yoshida
  • Shigenori Kanaya
  • Haruki Nakamura
Chapter
Part of the NATO ASI Series book series (NSSA, volume 225)

Abstract

The atomic scale investigation of proteins by NMR primarily depends on the assignment of the individual resonance peaks. The assignment strategy now utilized for smaller proteins (<10 kDa) consists of two steps1: 1) The identification of the spin system characterizing each of the amino acids by COSY and 2) The sequential connection of the identified spin systems along the amino acid sequence by NOESY. To extend the strategy to larger proteins, the overlapping of chemical shifts of the densely scattered resonance should be eliminated. A new strategy recently developed in our group utilizes heteronuclear 3D NMR2,3 and isotope enrichment to a large extent to solve the problem.

Keywords

Cross Peak Sequential Assignment Scattered Resonance HMQC Spectrum Full Assignment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    K. Wüthrich, “NMR of Proteins and Nucleic Acids”, John-Wiley, New York (1986).Google Scholar
  2. 2.
    K. Nagayama, T. Yamazaki, M. Yoshida, S. Kanaya, and H. Nakamura, J. Biochem. 108, 149–152 (1990).PubMedGoogle Scholar
  3. 3.
    T. Yamazaki, M. Yoshida, S. Kanaya, H. Nakamura, and K. Nagayama, Biochemistry, 30, 6036–6047 (1991).PubMedCrossRefGoogle Scholar
  4. 4.
    L. Muller, J. Am. Chem. Soc. 101, 4481–4482 (1977).CrossRefGoogle Scholar
  5. 5.
    D. Marion, et al., J. Am. Chem. Soc. 111, 1515–1517 (1989).CrossRefGoogle Scholar
  6. 6.
    S. Kanaya, A. Kohara, M. Miyagawa, T. Matsuzaki, K. Morikawa, and M. Ikehara, J. Biol. Chem. 264, 11546–11549 (1989).PubMedGoogle Scholar
  7. 7.
    K. Katayanagi, et al., Nature, 347, 306–309 (1990).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1991

Authors and Affiliations

  • Kuniaki Nagayama
    • 1
  • Toshio Yamazaki
    • 1
  • Mayumi Yoshida
    • 1
    • 2
  • Shigenori Kanaya
    • 1
    • 3
  • Haruki Nakamura
    • 1
    • 3
  1. 1.Biometrology LabJEOL Ltd.Musashino, Akishima, Tokyo 196Japan
  2. 2.Tokyo Research LaboratoriesKyowa Hakko, Machida, Tokyo 194Japan
  3. 3.Protein Engineering Research InstituteFuruedai, Suita, Osaka 565Japan

Personalised recommendations