[NMR paper] Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Related ArticlesSequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
Biochemistry. 2018 Feb 21;:
Authors: Alphonse S, Itin B, Khayat R, Ghose R
Abstract
While solid-state NMR (ssNMR) has emerged as a powerful technique to study viral capsids, current studies are limited to capsids formed from single proteins or single polyproteins. The ability to selectively label individual protein components within multi-protein viral capsids and the resulting spectral simplification will facilitate the extension of ssNMR techniques to complex viruses. In vitro capsid assembly by combining individually purified, labeled and unlabeled components in NMR quantities is not a viable option for most viruses. To overcome this barrier, we present a method that utilizes sequential protein expression and in cell assembly of component-specifically labeled viral capsids in amounts suitable for NMR studies. We apply this approach to purify capsids of bacteriophage 6 isotopically-labeled on only one of its four constituent protein components, the NTPase P4. Using P4-labeled 6 capsids and the sensitivity enhancement provided by dynamic nuclear polarization (DNP) we illustrate the utility of this method to enable ssNMR studies on complex viruses.
PMID: 29465229 [PubMed - as supplied by publisher]
Segmental isotopic labeling of HIV-1 capsid protein assemblies for solid state NMR
Segmental isotopic labeling of HIV-1 capsid protein assemblies for solid state NMR
Abstract
Recent studies of noncrystalline HIV-1 capsid protein (CA) assemblies by our laboratory and by Polenova and coworkers (Protein Sci 19:716â??730, 2010; J Mol Biol 426:1109â??1127, 2014; J Biol Chem 291:13098â??13112, 2016; J Am Chem Soc 138:8538â??8546, 2016; J Am Chem Soc 138:12029â??12032, 2016; J Am Chem Soc 134:6455â??6466, 2012; J Am Chem Soc 132:1976â??1987, 2010; J Am Chem Soc 135:17793â??17803, 2013; Proc Natl Acad Sci USA 112:14617â??14622, 2015; J Am...
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02-21-2018 12:45 AM
Structure of the Dimerization Interface in the Mature HIV-1 Capsid Protein Lattice from Solid State NMR of Tubular Assemblies
Structure of the Dimerization Interface in the Mature HIV-1 Capsid Protein Lattice from Solid State NMR of Tubular Assemblies
Marvin J. Bayro and Robert Tycko
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/jacs.6b03983/20160628/images/medium/ja-2016-03983c_0006.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.6b03983
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/yXorzqET3DQ
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06-29-2016 09:52 AM
[NMR paper] Structure of the dimerization interface in the mature HIV-1 capsid protein lattice from solid state NMR of tubular assemblies.
Structure of the dimerization interface in the mature HIV-1 capsid protein lattice from solid state NMR of tubular assemblies.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-pubmed-acspubs.jpg Related Articles Structure of the dimerization interface in the mature HIV-1 capsid protein lattice from solid state NMR of tubular assemblies.
J Am Chem Soc. 2016 Jun 14;
Authors: Bayro MJ, Tycko R
Abstract
The HIV-1 capsid protein (CA) forms the capsid shell that encloses RNA within a mature...
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06-15-2016 11:12 PM
[NMR paper] Sequential protein expression and selective labeling for in-cell NMR in human cells.
Sequential protein expression and selective labeling for in-cell NMR in human cells.
Related Articles Sequential protein expression and selective labeling for in-cell NMR in human cells.
Biochim Biophys Acta. 2015 Dec 23;
Authors: Luchinat E, Secci E, Cencetti F, Bruni P
Abstract
BACKGROUND: In-cell NMR is a powerful technique to investigate proteins in living human cells at atomic resolution. Ideally, when studying functional processes involving protein-protein interactions by NMR, only one partner should be isotopically...
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01-04-2016 07:49 PM
Sequential protein expression and selective labeling for in-cell NMR in human cells
Sequential protein expression and selective labeling for in-cell NMR in human cells
Publication date: Available online 23 December 2015
Source:Biochimica et Biophysica Acta (BBA) - General Subjects</br>
Author(s): Enrico Luchinat, Erica Secci, Francesca Cencetti, Paola Bruni</br>
Background In-cell NMR is a powerful technique to investigate proteins in living human cells at atomic resolution. Ideally, when studying functional processes involving protein-protein interactions by NMR, only one partner should be isotopically labeled. Here we show that...
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12-28-2015 12:26 AM
[NMR paper] Monitoring Binding of HIV-1 Capsid Assembly Inhibitors Using (19) F Ligand-and (15) N Protein-Based NMR and X-ray Crystallography: Early Hit Validation of a Benzodiazepine Series.
Monitoring Binding of HIV-1 Capsid Assembly Inhibitors Using (19) F Ligand-and (15) N Protein-Based NMR and X-ray Crystallography: Early Hit Validation of a Benzodiazepine Series.
Related Articles Monitoring Binding of HIV-1 Capsid Assembly Inhibitors Using (19) F Ligand-and (15) N Protein-Based NMR and X-ray Crystallography: Early Hit Validation of a Benzodiazepine Series.
ChemMedChem. 2013 Feb 10;
Authors: Goudreau N, Coulombe R, Faucher AM, Grand-Maître C, Lacoste JE, Lemke CT, Malenfant E, Bousquet Y, Fader L, Simoneau B, Mercier JF, Titolo S, Mason SW
...
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02-13-2013 12:47 PM
Ph.D. student in solid-state NMR: Studying protein folding and assembly at atomic sca
Ph.D. student in solid-state NMR: Studying protein folding and assembly at atomic scale
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The Faculty of Science consists of six departments: Biology, Pharmaceutical Sciences, Information and Computing Sciences, Physics and Astronomy, Mathematics and Chemistry. The Faculty is home to 3500 students and nearly 2000 staff and is internationally renowned for the quality of its research.
The NMR Research Group is part of the Chemistry Department (Universiteit Utrecht fac. Scheikunde) and belongs to the Bijvoet Center for Biomolecular...
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Cell-free expression and labeling strategies for a new decade in solid-state NMR.
Cell-free expression and labeling strategies for a new decade in solid-state NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Cell-free expression and labeling strategies for a new decade in solid-state NMR.
N Biotechnol. 2010 Aug 3;
Authors: Abdine A, Verhoeven MA, Warschawski DE
Although solid-state NMR and cell-free expression have recently become standard methods in biology, the combination of the two is still at a very early stage of development. In this...
Sequential Protein Expression and Capsid Assembly In Cell: Towards the Study of Multi-Protein Viral Capsids Using Solid-State NMR Techniques.
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