Journal of Biomolecular NMR

, Volume 2, Issue 1, pp 71–82 | Cite as

A1H-15N NM R study of human c-Ha-ras protein: Biosynthetic incorporation of15N-labeled amino acids

  • Kazuhiko Yamasaki
  • Yutaka Muto
  • Yutaka Ito
  • Markus Wälchli
  • Tatsuo Miyazawa
  • Susumu Nishimura
  • Shigeyuki Yokoyama
Research Papers

Summary

A1H-15N NMR study was performed on the GDP-bound form of a truncated human c-Ha-ras oncogene product (171 amino acid residues). Resonance cross peaks of the backbone amide1H-15N nuclei of a uniformly15N-labeled protein were observed with heteronuclear-single-quantum coherence spectroscopy (HSQC). In order to resolve overlapping cross peaks, selective15N-labeling of one or two types of amino acid residues (Ala, Arg, Asx, Glx, Gly, His, lie, Leu, Lys, Met, Phe, Ser; Thr, Tyr and/or Val) was carried out using appropriateE. coli mutant strains. By this procedure, all the backbone1H-15N cross peaks were classified into amino acid types.

Keywords

Selective15N-labeling Ras protein Amino acid type classification HMQC HSQC 

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Copyright information

© ESCOM Science Publishers B.V. 1992

Authors and Affiliations

  • Kazuhiko Yamasaki
    • 1
  • Yutaka Muto
    • 1
  • Yutaka Ito
    • 1
  • Markus Wälchli
    • 2
  • Tatsuo Miyazawa
    • 3
  • Susumu Nishimura
    • 4
  • Shigeyuki Yokoyama
    • 1
  1. 1.Department of Biophysics and Biochemistry, Faculty of ScienceUniversity of TokyoTokyoJapan
  2. 2.Bruker JapanIbarakiJapan
  3. 3.Faculty of EngineeringYokohama National UniversityYokohamaJapan
  4. 4.National Cancer Center Research InstituteTokyoJapan

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