A novel method of convection compensated ENMR (CC-ENMR) has been developed to detect electrophoretic motion of ionic species in the presence of bulk solution convection. This was accomplished using a gradient moment nulling technique to remove spectral artifacts from heat-induced convection and using the polarity switch of the applied electric field to retain spin phase modulations due to electrophoretic flow. Experiments were carried out with a mixture of 100 mM L-aspartic acid and 100 mM 4,9-dioxa-1,12-dodecanediamine to demonstrate this new method of ENMR. CC-ENMR enhances our previously developed capillary array ENMR (CA-ENMR) in solving the convection problem. The combined CA- and CC-ENMR approach strengthens the potential of multidimensional ENMR in simultaneous structural determination of coexisting proteins and protein conformations in biological buffer solutions of high ionic strength. Structural mapping of interacting proteins during biochemical reactions becomes possible in the future using ENMR techniques, which may have a profound impact on the understanding of biological events, including protein folding, genetic control, and signal transduction in general.
Simultaneous convection compensation and solvent suppression in biomolecular NMR diffusion experiments
Simultaneous convection compensation and solvent suppression in biomolecular NMR diffusion experiments
Abstract Thermal convection and high intensity solvent resonances can significantly hamper diffusion estimates in pulsed gradient spin-echo nuclear magnetic resonance diffusion experiments on biomolecule samples. To overcome these two problems, a new double functional NMR diffusion sequence, double echo PGSTE-WATERGATE, is presented. The new sequence provides excellent convection compensation and solvent suppression (with a suppression factor in excess of at least 105 in a single scan)...
nmrlearner
Journal club
0
01-09-2011 12:46 PM
[NMR paper] Trp repressor-operator binding: NMR and electrophoretic mobility shift studies of the
Trp repressor-operator binding: NMR and electrophoretic mobility shift studies of the effect of DNA sequence and corepressor binding on two Trp repressor-operator complexes.
Related Articles Trp repressor-operator binding: NMR and electrophoretic mobility shift studies of the effect of DNA sequence and corepressor binding on two Trp repressor-operator complexes.
Biochemistry. 2002 Dec 17;41(50):14866-78
Authors: Jaseja M, Jeeves M, Hyde EI
In Trp repressor-DNA complexes, most interactions either occur with phosphate groups or are...
nmrlearner
Journal club
0
11-24-2010 08:58 PM
[NMR paper] Constant-time multidimensional electrophoretic NMR.
Constant-time multidimensional electrophoretic NMR.
Related Articles Constant-time multidimensional electrophoretic NMR.
J Magn Reson. 2002 Jun;156(2):181-6
Authors: Li E, He Q
Multidimensional electrophoretic NMR (ENMR) has been introduced to determine structures of coexisting proteins and protein conformations in solution. Signals of different proteins are separated in a new dimension of electrophoretic flow according to their characteristic electrophoretic mobilities. The electrophoretic interferograms have been generated in the flow...
nmrlearner
Journal club
0
11-24-2010 08:49 PM
[NMR paper] Three-dimensional electrophoretic NMR correlation spectroscopy.
Three-dimensional electrophoretic NMR correlation spectroscopy.
Related Articles Three-dimensional electrophoretic NMR correlation spectroscopy.
J Magn Reson. 2000 Dec;147(2):361-5
Authors: He Q, Lin W, Liu Y, Li E
A novel method of three-dimensional electrophoretic NMR correlation spectroscopy (3D EP-COSY) has been proposed, developed, and implemented. It has a demonstrated potential of facilitating simultaneous structural assignments of multiple proteins in mixtures. The principle is to add a pulsed DC electric field that introduces a new...
nmrlearner
Journal club
0
11-19-2010 08:29 PM
[NMR paper] Capillary array electrophoretic NMR of proteins in biological buffer solutions.
Capillary array electrophoretic NMR of proteins in biological buffer solutions.
Related Articles Capillary array electrophoretic NMR of proteins in biological buffer solutions.
J Magn Reson. 1999 Dec;141(2):355-9
Authors: He Q, Liu Y, Sun H, Li E
The capillary array electrophoretic NMR (CA-ENMR) was developed to study protein mixtures in biological buffer solutions of high ionic strength. By enhancing the strength of the effective electric field across the sample, the technique permits the detection of the electrophoretic motion of 1 mM...
nmrlearner
Journal club
0
11-18-2010 08:31 PM
24-SEMA as a Sensitive and Offset Compensated SLF Sequence
24-SEMA as a Sensitive and Offset Compensated SLF Sequence
Publication year: 2010
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 3 September 2010</br>
S., Jayanthi , N., Sinha , K.V., Ramanathan</br>
Separated Local Field (SLF) spectroscopy is a powerful tool for the determination of structure and dynamics of oriented systems such as membrane proteins oriented in lipid bilayers and liquid crystals. Of many SLF techniques available, Polarization Inversion Spin Exchange at Magic Angle (PISEMA) has found wide application due to its many favorable...
Convection compensated electrophoretic NMR.
Linear Mode
