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Journal of Biomolecular NMR

, Volume 1, Issue 1, pp 13–22 | Cite as

Stereospecific assignment of β-methylene protons in larger proteins using 3D15N-separated Hartmann-Hahn and13C-separated rotating frame Overhauser spectroscopy

  • G. Marius Clore
  • Ad Bax
  • Angela M. Gronenborn
Research Papers

Summary

3Jxβ coupling constants and complementary nuclear Overhauser data on the intraresidue C x H−CβH distances form an essential part of the data needed to obtain stereospecific assignments of β-methylene protons in proteins. In this paper we show that information regarding the magnitude of the3Jxβ coupling constants can be extracted from a semi-quantitative interpretation of relative peak intensities in a 3D15N-separated1H−1H Hartmann-Hahn1H−15N multiple quantum coherence (HOHAHA-HMQC) spectrum. In addition, we demonstrate that reliable information on the intraresidue C x H−CβH distances, free of systematic errors arising from spin diffusion, can be obtained from a 3D13C-separated1H−1H rotating frame Overhauser effect1H−13C multiple quantum coherence (ROESY-HMQC) spectrum. The applicability of these experiments to larger proteins is illustrated with respect to interleukin-1β, a protein of 153 residues and 17.4 kDa molecular weight.

Keywords

Stereospecific assignment 3D NMR Heteronuclear NMR Hartmann-Hahn ROE 

Abbreviations

1L-1β

interleukin-1β

NOE

nuclear Overhauser effect

ROE

rotating frame Overhauser effect

HOHAHA

homonuclear Hartmann-Hahn spectroscopy

NOESY

nuclear Overhauser enhancement spectroscopy

ROESY

rotating frame Overhauser spectroscopy

HMQC

heteronuclear multiple quantum coherence spectroscopy

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Copyright information

© ESCOM Science Publishers B.V. 1991

Authors and Affiliations

  • G. Marius Clore
    • 1
  • Ad Bax
    • 1
  • Angela M. Gronenborn
    • 1
  1. 1.Laboratory of Chemical Physics, Building 2, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA

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