Related ArticlesDetergent Optimized Membrane Protein Reconstitution in Liposomes for Solid State NMR.
Biochemistry. 2014 Mar 25;
Authors: Murray DT, Griffin JM, Cross TA
Abstract
For small helical membrane proteins their structure is highly sensitive to their environment and solid state NMR is a structural technique that can characterize these membrane proteins in native like lipid bilayers and proteoliposomes. To date, a systematic method by which to evaluate the effect of the solubilizing detergent on proteoliposome preparations for solid state NMR of membrane proteins has not been presented in the literature. A set of experiments are presented aimed at determining the conditions most amenable to dialysis mediated reconstitution sample preparation. A membrane protein from M. tuberculosis is used to illustrate the method. The results show that a detergent that stabilizes the most protein is not always ideal and sometimes cannot be removed by dialysis. By focusing on the lipid and protein binding properties of the detergent, proteoliposome preparations can be readily produced which provide double signal the to noise ratios for both oriented sample and magic angle spinning solid state NMR. The method will allow more membrane protein drug targets to be structurally characterized in lipid bilayer environments.
PMID: 24665863 [PubMed - as supplied by publisher]
[NMR paper] Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy.
Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy.
Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy.
J Biomol NMR. 2013 Aug 21;
Authors: Mote KR, Gopinath T, Veglia G
Abstract
The low sensitivity inherent to both the static and magic angle spinning techniques of solid-state NMR (ssNMR) spectroscopy...
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08-23-2013 01:07 AM
[NMR paper] Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
Protein Expr Purif. 2013 Aug 1;
Authors: Bhate MP, Wylie BJ, Thompson A, Tian L, Nimigean C, McDermott AE
Abstract
We report the expression, purification, liposome reconstitution and functional validation of uniformly (13)C and (15)N isotope labeled...
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08-07-2013 01:24 PM
Micro-coil NMR to monitor optimization of the reconstitution conditions for the integral membrane protein OmpW in detergent micelles
Micro-coil NMR to monitor optimization of the reconstitution conditions for the integral membrane protein OmpW in detergent micelles
Abstract Optimization of aqueous solutions of the integral membrane protein (IMP) OmpW for NMR structure determination has been monitored with micro-coil NMR, which enables the acquisition of NMR spectra using only micrograms of protein and detergent. The detergent 30-Fos (2-undecylphosphocholine) was found to yield the best 2D -TROSY correlation NMR spectra of -labeled OmpW. For the OmpW structure determination we then optimized the 30-Fos concentration,...
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08-16-2012 09:18 AM
Investigation of the Interface in Silica-Encapsulated Liposomes by Combining Solid State NMR and First Principles Calculations
Investigation of the Interface in Silica-Encapsulated Liposomes by Combining Solid State NMR and First Principles Calculations
Nicolas Folliet, Claire Roiland, Sylvie Be?gu, Anne Aubert, Tzonka Mineva, Annick Goursot, Kaliaperumal Selvaraj, Luminita Duma, Frederik Tielens, Francesco Mauri, Guillaume Laurent, Christian Bonhomme, Christel Gervais, Florence Babonneau and Thierry Azai?s
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja201002r/aop/images/medium/ja-2011-01002r_0009.gif
Journal of the American Chemical Society
DOI:...
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10-05-2011 08:52 AM
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA.
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA.
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA.
J Am Chem Soc. 2011 Mar 1;
Authors: Renault M, Bos MP, Tommassen J, Baldus M
Multidomain proteins constitute a large part of prokaryotic and eukaryotic proteomes and play fundamental roles in various physiological processes. However, their structural characterization is challenging because of their large size and...
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03-03-2011 12:34 PM
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA
Marie Renault, Martine P. Bos, Jan Tommassen and Marc Baldus
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja109469c/aop/images/medium/ja-2010-09469c_0004.gif
Journal of the American Chemical Society
DOI: 10.1021/ja109469c
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/9XN1qiW-S-I
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03-02-2011 02:01 AM
[NMR paper] Bead-linked proteoliposomes: a reconstitution method for nmr analyses of membrane protein-ligand interactions.
Bead-linked proteoliposomes: a reconstitution method for nmr analyses of membrane protein-ligand interactions.
Related Articles Bead-linked proteoliposomes: a reconstitution method for nmr analyses of membrane protein-ligand interactions.
J Am Chem Soc. 2005 Aug 31;127(34):12021-7
Authors: Yokogawa M, Takeuchi K, Shimada I
Structural information about the interactions between membrane proteins and their ligands provides insights into the membrane protein functions. A variety of surfactants have been used for structural analyses of membrane...
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12-01-2010 06:56 PM
[NMR paper] Transverse dephasing optimized solid-state NMR spectroscopy.
Transverse dephasing optimized solid-state NMR spectroscopy.
Related Articles Transverse dephasing optimized solid-state NMR spectroscopy.
J Am Chem Soc. 2003 Nov 19;125(46):13938-9
Authors: De Paëpe G, Giraud N, Lesage A, Hodgkinson P, Böckmann A, Emsley L
It is shown how coherence lifetimes in solid-state NMR experiments can be controlled. New decoupling schemes are introduced which actively optimize dephasing times, providing increases of up to a factor of 2 with respect to the best existing schemes. The new schemes are implemented in...