NMR structure determination and investigation using a reduced proton (REDPRO) labeling strategy for proteins

Alexander Shekhtman, Ranajeet Ghose, Michael Goger, David Cowburn

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

We present here a stable isotope labeling technique for proteins, which seeks the appropriate compromise between the advantages of (a) random isotope labeling, with its large number of protons available for structure determination, and (b) selective labeling to generate isolated proton spins decreasing spectral complexity and improving relaxation properties of NMR experiments. The described reduced proton (REDPRO) procedure results in side-chain specific protonation of overexpressed proteins, which is highly selective. The REDPRO labeling scheme provides a sufficient number of NOE constraints for structure calculation. Dramatically improved relaxation properties of the heteronuclear magnetization transfer coupled with TROSY advantages make the proposed labeling scheme an attractive approach for study of high molecular weight protein targets, their ligand sites, and interdomain interactions.

Original languageEnglish (US)
Pages (from-to)177-182
Number of pages6
JournalFEBS Letters
Volume524
Issue number1-3
DOIs
StatePublished - Jul 31 2002
Externally publishedYes

Keywords

  • Nuclear magnetic resonance relaxation
  • Solution structure determination
  • Stable isotope labeling

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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