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

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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.

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Correspondence to Shigeyuki Yokoyama.

Additional information

This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan

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Yamasaki, K., Muto, Y., Ito, Y. et al. A1H-15N NM R study of human c-Ha-ras protein: Biosynthetic incorporation of15N-labeled amino acids. J Biomol NMR 2, 71–82 (1992). https://doi.org/10.1007/BF02192801

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Keywords

  • Selective15N-labeling
  • Ras protein
  • Amino acid type classification
  • HMQC
  • HSQC