Abstract Magic-angle spinning (MAS) solid-state NMR (SSNMR) spectroscopy of uniformly-13C,15N labeled protein samples provides insight into atomic-resolution chemistry and structure. Data collection efficiency has advanced remarkably in the last decade; however, the study of larger proteins is still challenged by relatively low resolution in comparison to solution NMR. In this study, we present a systematic analysis of SSNMR protein spectra acquired at 11.7, 17.6 and 21.1 Tesla (1H frequencies of 500, 750, and 900 MHz). For two protein systemsâ??GB1, a 6 kDa nanocrystalline protein and DsbA, a 21 kDa nanocrystalline proteinâ??line narrowing is demonstrated in all spectral regions with increasing field. Resolution enhancement is greatest in the aliphatic region, including methine, methylene and methyl sites. The resolution for GB1 increases markedly as a function of field, and for DsbA, resolution in the Câ??C region increases by 42%, according to the number of peaks that can be uniquely picked and integrated in the 900 MHz spectra when compared to the 500 MHz spectra. Additionally, chemical exchange is uniquely observed in the highest field spectra for at least two isoleucine Cδ1 sites in DsbA. These results further illustrate the benefits of high-field MAS SSNMR spectroscopy for protein structural studies.
Content Type Journal Article
Pages 149-155
DOI 10.1007/s10858-009-9389-9
Authors
Lindsay J. Sperling, University of Illinois at Urbana-Champaign Department of Chemistry 600 South Mathews Avenue Urbana IL 61801 USA
Andrew J. Nieuwkoop, University of Illinois at Urbana-Champaign Department of Chemistry 600 South Mathews Avenue Urbana IL 61801 USA
Andrew S. Lipton, Pacific Northwest National Laboratory Environmental Molecular Sciences Laboratory Richland WA 99352 USA
Deborah A. Berthold, University of Illinois at Urbana-Champaign Department of Chemistry 600 South Mathews Avenue Urbana IL 61801 USA
Chad M. Rienstra, University of Illinois at Urbana-Champaign Department of Chemistry 600 South Mathews Avenue Urbana IL 61801 USA
High dimensional and high resolution pulse sequences for backbone resonance assignment of intrinsically disordered proteins
High dimensional and high resolution pulse sequences for backbone resonance assignment of intrinsically disordered proteins
Abstract Four novel 5D (HACA(N)CONH, HNCOCACB, (HACA)CON(CA)CONH, (H)NCO(NCA)CONH), and one 6D ((H)NCO(N)CACONH) NMR pulse sequences are proposed. The new experiments employ non-uniform sampling that enables achieving high resolution in indirectly detected dimensions. The experiments facilitate resonance assignment of intrinsically disordered proteins. The novel pulse sequences were successfully tested using δ subunit (20 kDa) of Bacillus subtilis RNA polymerase...
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02-21-2012 03:40 AM
Ultra-high resolution in MAS solid-state NMR of perdeuterated proteins: Implications for Structure and Dynamics
Ultra-high resolution in MAS solid-state NMR of perdeuterated proteins: Implications for Structure and Dynamics
Publication year: 2012
Source: Journal of Magnetic Resonance, Available online 5 January 2012</br>
Bernd*Reif</br>
http://www.sciencedirect.com/cache/MiamiImageURL/1-s2.0-S1090780711005969-fx1.sml</br></br></br>
Source: Journal of Magnetic Resonance
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Ultra High-Resolution NMR: Sustained Induction Decays of Long-Lived Coherences
Ultra High-Resolution NMR: Sustained Induction Decays of Long-Lived Coherences
Aure?lien Bornet, Sami Jannin, J. A. (Ton) Konter, Patrick Hautle, Ben van den Brandt and Geoffrey Bodenhausen
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja2052792/aop/images/medium/ja-2011-052792_0005.gif
Journal of the American Chemical Society
DOI: 10.1021/ja2052792
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/F3VMiSJp4h4
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09-02-2011 07:31 PM
[NMR paper] High-resolution NMR spectroscopy of membrane proteins in aligned bicelles.
High-resolution NMR spectroscopy of membrane proteins in aligned bicelles.
Related Articles High-resolution NMR spectroscopy of membrane proteins in aligned bicelles.
J Am Chem Soc. 2004 Dec 1;126(47):15340-1
Authors: De Angelis AA, Nevzorov AA, Park SH, Howell SC, Mrse AA, Opella SJ
High-resolution solid-state NMR spectra can be obtained from uniformly (15)N-labeled membrane proteins in magnetically aligned bicelles. Fast uniaxial diffusion about the axis of the bilayer normal results in single-line spectra that contain the orientational...
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11-24-2010 10:03 PM
[NMR paper] Sensitive high resolution inverse detection NMR spectroscopy of proteins in the solid
Sensitive high resolution inverse detection NMR spectroscopy of proteins in the solid state.
Related Articles Sensitive high resolution inverse detection NMR spectroscopy of proteins in the solid state.
J Am Chem Soc. 2003 Dec 24;125(51):15831-6
Authors: Paulson EK, Morcombe CR, Gaponenko V, Dancheck B, Byrd RA, Zilm KW
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...
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11-24-2010 09:16 PM
[NMR paper] Ultra-high-field MAS NMR assay of a multispin labeled ligand bound to its G-protein r
Ultra-high-field MAS NMR assay of a multispin labeled ligand bound to its G-protein receptor target in the natural membrane environment: electronic structure of the retinylidene chromophore in rhodopsin.
Related Articles Ultra-high-field MAS NMR assay of a multispin labeled ligand bound to its G-protein receptor target in the natural membrane environment: electronic structure of the retinylidene chromophore in rhodopsin.
Biochemistry. 2001 Mar 20;40(11):3282-8
Authors: Verhoeven MA, Creemers AF, Bovee-Geurts PH, De Grip WJ, Lugtenburg J, de Groot HJ
...
High resolution NMR spectroscopy of nanocrystalline proteins at ultra-high magnetic field
Linear Mode
