(4,2)D Projection--reconstruction experiments for protein backbone assignment: application to human carbonic anhydrase II and calbindin D(28K).
Projection-reconstruction NMR experiments have been shown to significantly reduce the acquisition time required to obtain protein backbone assignment data. To date, this concept has only been applied to smaller (15)N/(13)C-labeled proteins. Here, we show that projection-reconstruction NMR techniques can be extended to larger protonated and perdeuterated proteins. We present a suite of (4,2)D triple-resonance experiments for protein backbone assignment and a Hybrid Backprojection/Lower-Value algorithm for reconstructing data with relatively weak signal-to-noise ratios. In addition, we propose a sampling theorem and discuss its implication on the choice of projection angles. We demonstrate the efficacy of this approach using the 29 kDa protein, human carbonic anhydrase II and the 30 kDa protein, calbindin D(28K).
Venters
Ronald A
RA
Duke University NMR Center, Duke University Medical Center, Durham, North Carolina 27710, USA. venters@windsurf.mc.duke.edu
Coggins
Brian E
BE
Kojetin
Doug
D
Cavanagh
John
J
Zhou
Pei
P
D064026:Calbindins; D024402:Carbonic Anhydrase II / Q000737:chemistry*; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D064030:S100 Calcium Binding Protein G / Q000737:chemistry*
D064026:Calbindins; D064030:S100 Calcium Binding Protein G; D024402:Carbonic Anhydrase II