SOFAST-HMQC experiments for recording two-dimensional heteronuclear correlation spectra of proteins within a few seconds

J Biomol NMR. 2005 Dec;33(4):199-211. doi: 10.1007/s10858-005-4425-x.

Abstract

Fast multidimensional NMR with a time resolution of a few seconds provides a new tool for high throughput screening and site-resolved real-time studies of kinetic molecular processes by NMR. Recently we have demonstrated the feasibility to record protein 1H-15N correlation spectra in a few seconds of acquisition time using a new SOFAST-HMQC experiment (Schanda and Brutscher (2005) J. Am. Chem. Soc. 127, 8014). Here, we investigate in detail the performance of SOFAST-HMQC to record 1H-15N and 1H-13C correlation spectra of proteins of different size and at different magnetic field strengths. Compared to standard 1H-15N correlation experiments SOFAST-HMQC provides a significant gain in sensitivity, especially for fast repetition rates. Guidelines are provided on how to set up SOFAST-HMQC experiments for a given protein sample. In addition, an alternative pulse scheme, IPAP-SOFAST-HMQC is presented that allows application on NMR spectrometers equipped with cryogenic probes, and fast measurement of one-bond 1H-13C and 1H-15N scalar and residual dipolar coupling constants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Hydrogen
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Oxidoreductases / chemistry
  • Proteins / chemistry*
  • Ubiquitin / chemistry

Substances

  • Nitrogen Isotopes
  • Proteins
  • Ubiquitin
  • Hydrogen
  • Oxidoreductases