title: SOFAST-HMQC experiments for recording two-dimensional heteronuclear correlation spectra of proteins within a few seconds.
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.
authors.affiliation: Institut de Biologie Structurale-Jean-Pierre Ebel, UMR5075 CNRS-CEA-UJF, 41, rue Jules Horowitz, 38027, Grenoble Cedex, France.
authors.forename: Paul
authors.initials: P
authors.lastname: Schanda
authors.forename: Eriks
authors.initials: E
authors.lastname: Kupce
authors.forename: Bernhard
authors.initials: B
authors.lastname: Brutscher
mesh_terms: D006859:Hydrogen; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010088:Oxidoreductases / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry
chemical_list: D009587:Nitrogen Isotopes; D011506:Proteins; D025801:Ubiquitin; D006859:Hydrogen; D010088:Oxidoreductases