Sensitive High Resolution Inverse Detection NMR Spectroscopy of Proteins in the Solid State

Citations Over TimeTop 10% of 2003 papers

Abstract

A new indirect detection scheme for obtaining (15)N/(1)H shift correlation spectra in crystalline proteins is described. Excellent water suppression is achieved without the need for pulsed field gradients, and using only a 2-step phase cycle. Careful attention to overall NMR instrument stability was found critical for obtaining the best resolution and sensitivity. Magnetic dilution by deuteration of the protein in combination with high-speed magic angle spinning produces (1)H resonances averaging only 0.22 ppm in width, and in some cases lines as narrow as 0.17 ppm are obtained. In application to two different polymorphs of ubiquitin, structure dependent differences in both (15)N and (1)H amide chemical shifts are observed. In one case, distinct shifts for different molecules in the asymmetric unit are seen, and all differ substantially from solution NMR shifts. A gain of 7 in sensitivity makes the method competitive with solution NMR as long as nanocrystalline samples are available.

Related Papers

• → 27Al Solid-state NMR Structural Studies of Hydrotalcite Compounds Calcined at Different Temperatures(2009)21 cited
• → Predicting15N amide chemical shifts in proteins. I. An additive model for the backbone contribution(2001)12 cited
• → Recent developments in solid–state magic–angle spinning, nuclear magnetic resonance of fully and significantly isotopically labelled peptides and proteins(2004)16 cited
• → Characterization of V2O5-AlPO4 catalysts by 51V and 1H magic-angle spinning solid-state nuclear magnetic resonance spectroscopy(1995)7 cited
• → Magic‐Angle Spinning Solid‐State Nuclear Magnetic Resonance: Application to Structural Biology(2009)2 cited