UltraSOFAST HMQC NMR and the repetitive acquisition of 2D protein spectra at Hz rates.
Following unidirectional biophysical events such as the folding of proteins or the equilibration of binding interactions, requires experimental methods that yield information at both atomic-level resolution and at high repetition rates. Toward this end a number of different approaches enabling the rapid acquisition of 2D NMR spectra have been recently introduced, including spatially encoded "ultrafast" 2D NMR spectroscopy and SOFAST HMQC NMR. Whereas the former accelerates acquisitions by reducing the number of scans that are necessary for completing arbitrary 2D NMR experiments, the latter operates by reducing the delay between consecutive scans while preserving sensitivity. Given the complementarities between these two approaches it seems natural to combine them into a single tool, enabling the acquisition of full 2D protein NMR spectra at high repetition rates. We demonstrate here this capability with the introduction of "ultraSOFAST" HMQC NMR, a spatially encoded and relaxation-optimized approach that can provide 2D protein correlation spectra at approximately 1 s repetition rates for samples in the approximately 2 mM concentration range. The principles, relative advantages, and current limitations of this new approach are discussed, and its application is exemplified with a study of the fast hydrogen-deuterium exchange characterizing amide sites in Ubiquitin.
Gal
Maayan
M
Department of Chemical Physics, Weizmann Institute of Science, 76100 Rehovot, Israel.
Schanda
Paul
P
Brutscher
Bernhard
B
Frydman
Lucio
L
D000577:Amides / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D017510:Protein Folding; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013997:Time Factors; D025801:Ubiquitin / Q000032:analysis / Q000737:chemistry
D000577:Amides; D011506:Proteins; D025801:Ubiquitin; D006859:Hydrogen; D003903:Deuterium