Aromatic amino-acid side chains are essential components for the structure and function of proteins. We present herein a set of NMR experiments for time-efficient resonance assignment of histidine and tyrosine side chains in uniformly 13C/15N-labeled proteins. The use of band-selective 13C pulses allows to deal with linear chains of coupled spins, thus avoiding signal loss that occurs in branched spin systems during coherence transfer. Furthermore, our pulse schemes make use of longitudinal 1H relaxation enhancement, Ernst-angle excitation, and simultaneous detection of 1H and 13C steady-state polarization to achieve significant signal enhancements.
15 N and 13 C- SOFAST-HMQC editing enhances 3D-NOESY sensitivity in highly deuterated, selectively [ 1 H, 13 C]-labeled proteins
15 N and 13 C- SOFAST-HMQC editing enhances 3D-NOESY sensitivity in highly deuterated, selectively -labeled proteins
Abstract
The ongoing NMR method development effort strives for high quality multidimensional data with reduced collection time. Here, we apply â??SOFAST-HMQCâ?? to frequency editing in 3D NOESY experiments and demonstrate the sensitivity benefits using highly deuterated and 15N, methyl labeled samples in H2O. The experiments benefit from a combination of selective T 1 relaxation (or L-optimized effect), from...
nmrlearner
Journal club
0
11-25-2016 02:22 AM
[NMR paper] NMR Scalar Couplings Across Intermolecular Hydrogen Bonds Between Zinc-Finger Histidine Side Chains and DNA Phosphate Groups.
NMR Scalar Couplings Across Intermolecular Hydrogen Bonds Between Zinc-Finger Histidine Side Chains and DNA Phosphate Groups.
NMR Scalar Couplings Across Intermolecular Hydrogen Bonds Between Zinc-Finger Histidine Side Chains and DNA Phosphate Groups.
J Phys Chem B. 2016 Sep 29;
Authors: Chattopadhyay A, Esadze A, Roy S, Iwahara J
Abstract
NMR scalar couplings across hydrogen bonds represent direct evidence of the partial covalent nature of hydrogen bonds and provide structural and dynamic information on hydrogen bonding. In...
nmrlearner
Journal club
0
09-30-2016 10:11 AM
Time-shared experiments for efficient assignment of triple-selectively labeled proteins
Time-shared experiments for efficient assignment of triple-selectively labeled proteins
Publication date: Available online 30 September 2014
Source:Journal of Magnetic Resonance</br>
Author(s): Frank Löhr , Aisha Laguerre , Christoph Bock , Sina Reckel , Peter J. Connolly , Norzehan Abdul-Manan , Franz Tumulka , Rupert Abele , Jonathan M. Moore , Volker Dötsch</br>
Combinatorial triple-selective labeling facilitates the NMR assignment process for proteins that are subject to signal overlap and insufficient signal-to-noise in standard triple-resonance...
nmrlearner
Journal club
0
10-01-2014 02:48 AM
[NMR paper] Improved NMR experiments with (13)C-isotropic mixing for assignment of aromatic and aliphatic side chains in labeled proteins.
Improved NMR experiments with (13)C-isotropic mixing for assignment of aromatic and aliphatic side chains in labeled proteins.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--production.springer.de-OnlineResources-Logos-springerlink.gif Related Articles Improved NMR experiments with (13)C-isotropic mixing for assignment of aromatic and aliphatic side chains in labeled proteins.
J Biomol NMR. 2014 Jan 4;
Authors: Kovacs H, Gossert A
Abstract
Three improved (13)C-spinlock experiments for side chain assignments of...
nmrlearner
Journal club
0
01-07-2014 11:16 PM
Multidimensional oriented solid-state NMR experiments enable the sequential assignment of uniformly 15N labeled integral membrane proteins in magnetically aligned lipid bilayers
Multidimensional oriented solid-state NMR experiments enable the sequential assignment of uniformly 15N labeled integral membrane proteins in magnetically aligned lipid bilayers
Abstract Oriented solid-state NMR is the most direct methodology to obtain the orientation of membrane proteins with respect to the lipid bilayer. The method consists of measuring 1H-15N dipolar couplings (DC) and 15N anisotropic chemical shifts (CSA) for membrane proteins that are uniformly aligned with respect to the membrane bilayer. A significant advantage of this approach is that tilt and azimuthal...
nmrlearner
Journal club
0
10-10-2011 06:27 AM
4D APSY-HBCB(CG)CDHD experiment for automated assignment of aromatic amino acid side chains in proteins
4D APSY-HBCB(CG)CDHD experiment for automated assignment of aromatic amino acid side chains in proteins
Abstract A four-dimensional (4D) APSY (automated projection spectroscopy)-HBCB(CG)CDHD experiment is presented. This 4D experiment correlates aromatic with aliphatic carbon and proton resonances from the same amino acid side chain of proteins in aqueous solution. It thus allows unambiguous sequence-specific assignment of aromatic amino acid ring signals based on backbone assignments. Compared to conventional 2D approaches, the inclusion of evolution periods on 1Hβ and 13Cδ...
nmrlearner
Journal club
0
09-30-2011 08:01 PM
[NMR paper] NMR assignment of protein side chains using residue-correlated labeling and NOE spect
NMR assignment of protein side chains using residue-correlated labeling and NOE spectra.
Related Articles NMR assignment of protein side chains using residue-correlated labeling and NOE spectra.
J Magn Reson. 2003 Dec;165(2):237-47
Authors: Mueller GA, Kirby TW, DeRose EF, London RE
A new approach for the isotopic labeling of proteins is proposed that aims to facilitate side chain resonance assignments. Residue-correlated (RC) labeling is achieved by the expression of a protein on a medium containing a mixture of labeled, e.g., amino acids,...
nmrlearner
Journal club
0
11-24-2010 09:16 PM
[NMR paper] Determination of pKa values of the histidine side chains of phosphatidylinositol-spec
Determination of pKa values of the histidine side chains of phosphatidylinositol-specific phospholipase C from Bacillus cereus by NMR spectroscopy and site-directed mutagenesis.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Determination of pKa values of the histidine side chains of phosphatidylinositol-specific phospholipase C from Bacillus cereus...
BEST and SOFAST experiments for resonance assignment of histidine and tyrosine side chains in 13 C/ 15 N labeled proteins
Linear Mode
