Abstract We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record 13C- and 15N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that provide cross-peaks in the indirect dimension with low resolution due to limits in experimental time. The technique is particularly advantageous at high field where even longer experimental times would be required for comparable resolution in NOESY-HSQC experiments. The method is demonstrated at 900 MHz and at 750 MHz on 37 and 31 kDa proteins, respectively. The resolution and time saving provided in this experiment was crucial for solving the structures of these two proteins.
Content Type Journal Article
Pages 311-318
DOI 10.1007/s10858-009-9372-5
Authors
Dominique P. Frueh, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology 240 Longwood Avenue Boston MA 02115 USA
Alison Leed, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology 240 Longwood Avenue Boston MA 02115 USA
Haribabu Arthanari, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology 240 Longwood Avenue Boston MA 02115 USA
Alexander Koglin, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology 240 Longwood Avenue Boston MA 02115 USA
Christopher T. Walsh, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology 240 Longwood Avenue Boston MA 02115 USA
Gerhard Wagner, Harvard Medical School Department of Biological Chemistry and Molecular Pharmacology 240 Longwood Avenue Boston MA 02115 USA
[NMR Sparky Yahoo group] R: [nmr_sparky] Re: distance constraints from NOESY crosspeaks
R: Re: distance constraints from NOESY crosspeaks
A single spectrum at 300 ms contains a lot of spin diffusion. It's good to assign DNA/RNA not for constraints. Try to use something like 50 ms instead. Marco
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[NMR Sparky Yahoo group] Re: distance constraints from NOESY crosspeaks
Re: distance constraints from NOESY crosspeaks
Did you do the build up with different mixing time?Jay From: jennig148 To: nmr_sparky@yahoogroups.com Sent: Friday, February 10,
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nmrlearner
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02-10-2012 09:28 PM
[NMR Sparky Yahoo group] distance constraints from NOESY crosspeaks
distance constraints from NOESY crosspeaks
Hello users, I have a NOESY spectrum of a DNA/RNA strand. I try to get the distance constraints by integration of the NOESY crosspeaks and then converting the
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nmrlearner
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02-10-2012 09:28 PM
High-resolution NMR field-cycling device for full-range relaxation and structural studies of biopolymers on a shared commercial instrument
High-resolution NMR field-cycling device for full-range relaxation and structural studies of biopolymers on a shared commercial instrument
Abstract Improvements are described in a shuttling field-cycling device (Redfield in Magn Reson Chem 41:753â??768, 2003), designed to allow widespread access to this useful technique by configuring it as a removable module to a commercial 500 MHz NMR instrument. The main improvements described here, leading to greater versatility, high reliability and simple construction, include: shuttling provided by a linear motor driven by an integrated-control...
nmrlearner
Journal club
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12-31-2011 10:40 AM
Automated sequence- and stereo-specific assignment of methyl-labeled proteins by paramagnetic relaxation and methylâ??methyl nuclear overhauser enhancement spectroscopy
Automated sequence- and stereo-specific assignment of methyl-labeled proteins by paramagnetic relaxation and methylâ??methyl nuclear overhauser enhancement spectroscopy
Abstract Methyl-transverse relaxation optimized spectroscopy is rapidly becoming the preferred NMR technique for probing structure and dynamics of very large proteins up to ~1 MDa in molecular size. Data interpretation, however, necessitates assignment of methyl groups which still presents a very challenging and time-consuming process. Here we demonstrate that, in combination with a known 3D structure, paramagnetic...
nmrlearner
Journal club
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09-26-2011 06:42 AM
[NMR paper] Hydrodynamic radii of native and denatured proteins measured by pulse field gradient
Hydrodynamic radii of native and denatured proteins measured by pulse field gradient NMR techniques.
Related Articles Hydrodynamic radii of native and denatured proteins measured by pulse field gradient NMR techniques.
Biochemistry. 1999 Dec 14;38(50):16424-31
Authors: Wilkins DK, Grimshaw SB, Receveur V, Dobson CM, Jones JA, Smith LJ
Pulse field gradient NMR methods have been used to determine the effective hydrodynamic radii of a range of native and nonnative protein conformations. From these experimental data, empirical relationships...
nmrlearner
Journal club
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11-18-2010 08:31 PM
Simultaneous detection of amide and methyl correlations using a time shared NMR experiment: application to binding epitope mapping
Simultaneous detection of amide and methyl correlations using a time shared NMR experiment: application to binding epitope mapping
Peter Würtz, Olli Aitio, Maarit Hellman and Perttu Permi
Journal of Biomolecular NMR; 2007; 39(2) pp 97 - 105
Abstract:
Simultaneous recording of different NMR parameters is an efficient way to reduce the overall experimental time and speed up structural studies of biological macromolecules. This can especially be beneficial in the case of fast NMR-based drug screening applications or for collecting NOE restraints, where prohibitively long data collection...
Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins
Linear Mode
