Abstract An experiment is presented to determine 3JHNHα coupling constants, with significant advantages for applications to unfolded proteins. The determination of coupling constants for the peptide chain using 1D 1H, or 2D and 3D 1H-15N correlation spectroscopy is often hampered by extensive resonance overlap when dealing with flexible, disordered proteins. In the experiment detailed here, the overlap problem is largely circumvented by recording 1H-13C� correlation spectra, which demonstrate superior resolution for unfolded proteins. J-coupling constants are extracted from the peak intensities in a pair of 2D spin-echo difference experiments, affording rapid acquisition of the coupling data. In an application to the cytoplasmic domain of human neuroligin-3 (hNlg3cyt) data were obtained for 78 residues, compared to 54 coupling constants obtained from a 3D HNHA experiment. The coupling constants suggest that hNlg3cyt is intrinsically disordered, with little propensity for structure.
Content Type Journal Article
Pages 343-349
DOI 10.1007/s10858-009-9382-3
Authors
Renee Otten, University of Groningen Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute Nijenborgh 4 9747 AG Groningen The Netherlands
Kathleen Wood, University of Groningen Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute Nijenborgh 4 9747 AG Groningen The Netherlands
Frans A. A. Mulder, University of Groningen Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute Nijenborgh 4 9747 AG Groningen The Netherlands
Spin Echo Diagram.png
http://upload.wikimedia.org/wikimedia/commons/thumb/b/b2/Spin_Echo_Diagram.png/300px-Spin_Echo_Diagram.png
Uploaded by user "4lex" on Tue, 07 Sep 2010 17:13:00 UTC
Added to category on Mon, 07 Feb 2011 23:43:04 UTC
Original image: 2757×1317 pixel; 862.155 bytes.
Licensing : CC-BY-SA
Spin Echo Diagram.png
More...
nmrlearner
NMR pictures
0
02-08-2011 08:16 PM
[NMR paper] Comparison of X-ray and NMR structures: is there a systematic difference in residue contacts between X-ray- and NMR-resolved protein structures?
Comparison of X-ray and NMR structures: is there a systematic difference in residue contacts between X-ray- and NMR-resolved protein structures?
Related Articles Comparison of X-ray and NMR structures: is there a systematic difference in residue contacts between X-ray- and NMR-resolved protein structures?
Proteins. 2005 Jul 1;60(1):139-47
Authors: Garbuzynskiy SO, Melnik BS, Lobanov MY, Finkelstein AV, Galzitskaya OV
We have compared structures of 78 proteins determined by both NMR and X-ray methods. It is shown that X-ray and NMR structures...
nmrlearner
Journal club
0
12-01-2010 06:56 PM
[NMR paper] NMR difference spectroscopy with a dual saddle-coil difference probe.
NMR difference spectroscopy with a dual saddle-coil difference probe.
Related Articles NMR difference spectroscopy with a dual saddle-coil difference probe.
Anal Bioanal Chem. 2004 Mar;378(6):1520-7
Authors: Macnaughtan MA, Smith AP, Goldsbrough PB, Santini RE, Raftery D
A new difference probe for nuclear magnetic resonance (NMR) spectroscopy is presented. The difference probe uses two saddle-shaped coils to excite and detect two samples simultaneously. The samples are held in a specially modified 3-mm NMR tube with an Ultem plastic disk to...
nmrlearner
Journal club
0
11-24-2010 09:25 PM
[NMR paper] Probing residual interactions in unfolded protein states using NMR spin relaxation te
Probing residual interactions in unfolded protein states using NMR spin relaxation techniques: an application to delta131delta.
Related Articles Probing residual interactions in unfolded protein states using NMR spin relaxation techniques: an application to delta131delta.
J Am Chem Soc. 2003 Oct 1;125(39):11988-92
Authors: Choy WY, Kay LE
Residual interactions in delta131delta, a large disordered fragment of staphylococcal nuclease, have been probed at two different pHs using backbone (15)N and side-chain methyl (2)H NMR spin relaxation...
nmrlearner
Journal club
0
11-24-2010 09:16 PM
[NMR paper] NMR spin-echo studies of hydration properties of the molecular chaperone alpha-crysta
NMR spin-echo studies of hydration properties of the molecular chaperone alpha-crystallin in the bovine lens.
Related Articles NMR spin-echo studies of hydration properties of the molecular chaperone alpha-crystallin in the bovine lens.
Biochim Biophys Acta. 2002 Jul 29;1598(1-2):46-54
Authors: Babizhayev MA, Nikolayev GN, Goryachev SN, Bours J
The water-binding properties of bovine lens alpha-crystallin, collagen from calf skin and bovine serum albumin (BSA), were investigated with various techniques. The water absorptive capacity was...
nmrlearner
Journal club
0
11-24-2010 08:58 PM
[NMR paper] Improved spin-echo-edited NMR diffusion measurements.
Improved spin-echo-edited NMR diffusion measurements.
Related Articles Improved spin-echo-edited NMR diffusion measurements.
J Magn Reson. 2001 Dec;153(2):273-6
Authors: Otto WH, Larive CK
The need for simple and robust schemes for the analysis of ligand-protein binding has resulted in the development of diffusion-based NMR techniques that can be used to assay binding in protein solutions containing a mixture of several ligands. As a means of gaining spectral selectivity in NMR diffusion measurements, a simple experiment, the gradient modified...
nmrlearner
Journal club
0
11-19-2010 08:44 PM
[NMR paper] Determination of local protein structure by spin label difference 2D NMR: the region
Determination of local protein structure by spin label difference 2D NMR: the region neighboring Asp61 of subunit c of the F1F0 ATP synthase.
Related Articles Determination of local protein structure by spin label difference 2D NMR: the region neighboring Asp61 of subunit c of the F1F0 ATP synthase.
Biochemistry. 1995 Feb 7;34(5):1635-45
Authors: Girvin ME, Fillingame RH
Purified subunit c from the H(+)-transporting F1F0 ATP synthase of Escherichia coli folds as an antiparallel pair of extended helices in a solution of...
nmrlearner
Journal club
0
08-22-2010 03:41 AM
Self-diffusion in Polymer Systems studied by Magnetic Field-Gradient Spin-Echo NMR Me
Self-diffusion in Polymer Systems studied by Magnetic Field-Gradient Spin-Echo NMR Methods
Publication year: 2010
Source: Progress in Nuclear Magnetic Resonance Spectroscopy, In Press, Accepted Manuscript, Available online 13 April 2010</br>
Harald, Walderhaug , Olle, Söderman , Daniel, Topgaard</br>
More...
Comprehensive determination of 3JHNHα for unfolded proteins using 13C�-resolved spin-echo difference spectroscopy
Linear Mode
