Backbone conformational constraints in a microcrystalline U-15N-labeled protein by 3D dipolar-shift solid-state NMR spectroscopy

Journal of the American Chemical Society
W Trent FranksChad M Rienstra

Abstract

Structural studies of uniformly labeled proteins by magic-angle spinning NMR spectroscopy have rapidly matured in recent years. Site-specific chemical shifts of several proteins have been assigned and structures determined from 2D or 3D data sets containing internuclear distance information. Here we demonstrate the application of a complementary technique for constraining protein backbone geometry using a site-resolved 3D dipolar-shift pulse sequence. The dipolar line shapes report on the relative orientations of 1H-15N[i] to 1H-15N[i+1] dipole vectors, constraining the torsion angles phi[i] and psi[i]. In addition, from the same 3D data set, several 1H-15N[i] to1H-15N[i+2] line shapes are extracted to constrain the torsion angles phi[i], psi[i], phi[i+1], and psi[i+1]. We report results for the majority of sites in the 56-residue beta1 immunoglobulin binding domain of protein G (GB1), using 3D experiments at 600 MHz 1H frequency. Excellent agreement between the SSNMR results and a new 1.14 A crystal structure illustrate the general potential of this technique for high-resolution structural refinement of solid proteins.

References

Jun 30, 2000·Journal of Magnetic Resonance·B ReifR G Griffin
Aug 1, 2002·Proceedings of the National Academy of Sciences of the United States of America·Chad M RienstraRobert G Griffin
Oct 7, 2004·Current Opinion in Structural Biology·Ann E McDermott
Mar 4, 2005·Angewandte Chemie·Li YongMartin Kaupp

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Citations

Aug 10, 2007·Journal of Biomolecular NMR·W Trent FranksChad M Rienstra
Aug 31, 2013·Journal of Biomolecular NMR·Matthew T EddyJudith Herzfeld
May 15, 2010·Journal of the American Chemical Society·Andrew J Nieuwkoop, Chad M Rienstra
Apr 2, 2009·Journal of the American Chemical Society·Józef R LewandowskiRobert G Griffin
Dec 7, 2007·The Journal of Physical Chemistry. B·Heather L Frericks SchmidtChad M Rienstra
Apr 24, 2012·Nature Chemistry·Ishita SenguptaChristopher P Jaroniec
Mar 18, 2008·Proceedings of the National Academy of Sciences of the United States of America·W Trent FranksChad M Rienstra
Feb 13, 2008·The Journal of Chemical Physics·Benjamin J Wylie, Chad M Rienstra
Sep 18, 2012·The Journal of Chemical Physics·Wei Qiang, Robert Tycko
Mar 12, 2013·Journal of Magnetic Resonance·Jun Li, Patrick C A van der Wel
Apr 28, 2017·Journal of the American Chemical Society·Kevin J DonovanRobert G Griffin
Dec 17, 2008·Structure·Paul RobustelliMichele Vendruscolo
Feb 24, 2009·Solid State Nuclear Magnetic Resonance·Karsten SeidelMarc Baldus
Jun 26, 2007·Protein Science : a Publication of the Protein Society·Tomoo OhashiHarold P Erickson
Dec 25, 2007·Magnetic Resonance in Chemistry : MRC·Seth A McNeillJoanna R Long
Dec 18, 2007·Magnetic Resonance in Chemistry : MRC·Anja Böckmann
Jan 5, 2008·Physical Chemistry Chemical Physics : PCCP·Benjamin J WylieChad M Rienstra
May 29, 2010·Journal of the American Chemical Society·Józef R LewandowskiLyndon Emsley
Dec 6, 2012·The Journal of Physical Chemistry Letters·Luca MollicaMartin Blackledge
Sep 29, 2007·Journal of Magnetic Resonance·Amir GoldbourtAnn E McDermott
Jan 29, 2008·Journal of the American Chemical Society·Manish A MehtaJoanna R Long

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