A 1H-15N NMR study of human c-Ha-ras protein: biosynthetic incorporation of 15N-labeled amino acids

J Biomol NMR. 1992 Jan;2(1):71-82. doi: 10.1007/BF02192801.

Abstract

A 1H-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 amide 1H-15N nuclei of a uniformly 15N-labeled protein were observed with heteronuclear single-quantum coherence spectroscopy (HSQC). In order to resolve overlapping cross peaks, selective 15N-labeling of one or two types of amino acid residues (Ala, Arg, Asx, Glx, Gly, His, Ile, Leu, Lys, Met, Phe, Ser, Thr, Tyr and/or Val) was carried out using appropriate E. coli mutant strains. By this procedure, all the backbone 1H-15N cross peaks were classified into amino acid types.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / metabolism
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Genes, Synthetic
  • Genes, ras
  • Humans
  • Hydrogen
  • Isotope Labeling / methods
  • Magnetic Resonance Spectroscopy / methods
  • Nitrogen Isotopes
  • Protein Conformation
  • Proto-Oncogene Proteins p21(ras) / biosynthesis
  • Proto-Oncogene Proteins p21(ras) / chemistry*
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics

Substances

  • Amino Acids
  • Nitrogen Isotopes
  • Recombinant Proteins
  • Hydrogen
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)