The non structural protein 5A (NS5A) regulates the replication of the hepatitis C viral RNA through a direct molecular interaction of its domain 2 (NS5A-D2) with the RNA dependent RNA polymerase NS5B. Because of conflicting data in the literature, we study here this molecular interaction using fluorinated versions of the NS5A-D2 protein derived from the JFH1 Hepatitis C Virus strain. Two methods to prepare fluorine-labelled NS5A-D2 involving the biosynthetic incorporation of a 19F-tryptophan using 5-fluoroindole and the posttranslational introduction of fluorine by chemical conjugation of 2-iodo-N-(trifluoromethyl)acetamide with the NS5A-D2 cysteine side chains are presented. The dissociation constants (KD) between NS5A-D2 and NS5B obtained with these two methods are in good agreement, and yield values comparable to those derived previously from a surface plasmon resonance study. We compare benefits and limitations of both labeling methods to study the interaction between an intrinsically disordered protein and a large molecular target by 19F NMR.
[NMR paper] NMR reveals the intrinsically disordered domain 2 of NS5A protein as an allosteric regulator of the hepatitis C virus RNA polymerase NS5B.
NMR reveals the intrinsically disordered domain 2 of NS5A protein as an allosteric regulator of the hepatitis C virus RNA polymerase NS5B.
Related Articles NMR reveals the intrinsically disordered domain 2 of NS5A protein as an allosteric regulator of the hepatitis C virus RNA polymerase NS5B.
J Biol Chem. 2017 Sep 14;:
Authors: Bessa LM, Launay H, Dujardin M, Cantrelle FX, Lippens G, Landrieu I, Schneider R, Hanoulle X
Abstract
Non-structural protein 5B (NS5B) is the RNAdependent RNA polymerase that catalyses replication of the...
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Overall Structural Model of NS5A Protein from HepatitisC Virus and Modulation by Mutations Confering Resistance of VirusReplication to Cyclosporin A
Overall Structural Model of NS5A Protein from HepatitisC Virus and Modulation by Mutations Confering Resistance of VirusReplication to Cyclosporin A
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.7b00212/20170607/images/medium/bi-2017-00212h_0013.gif
Biochemistry
DOI: 10.1021/acs.biochem.7b00212
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[NMR paper] The casein kinase 2-dependent phosphorylation of NS5A domain 3 from hepatitis C virus followed by time-resolved NMR.
The casein kinase 2-dependent phosphorylation of NS5A domain 3 from hepatitis C virus followed by time-resolved NMR.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--media.wiley.com-assets-7315-19-Wiley_FullText_120x30_orange.png http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--media.wiley.com-assets-7315-19-Wiley_FullText_120x30_orange.png Related Articles The casein kinase 2-dependent phosphorylation of NS5A domain 3 from hepatitis C virus followed by time-resolved NMR.
Chembiochem. 2015 Dec 18;
...
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(13)C/(15)N-(19)F Intermolecular REDOR NMR Study of the Interaction of TAR RNA with T
(13)C/(15)N-(19)F Intermolecular REDOR NMR Study of the Interaction of TAR RNA with Tat Peptides.
(13)C/(15)N-(19)F Intermolecular REDOR NMR Study of the Interaction of TAR RNA with Tat Peptides.
J Am Chem Soc. 2010 Nov 24;
Authors: Huang W, Varani G, Drobny GP
The complex of the HIV TAR RNA with the viral regulatory protein Tat is of considerable interest, but the plasticity of this interaction has made it impossible so far to establish the structure of that complex. In order to explore a new approach to obtain structural information on...
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13C/15N-19F Intermolecular REDOR NMR Study of the Interaction of TAR RNA with Tat Pep
13C/15N-19F Intermolecular REDOR NMR Study of the Interaction of TAR RNA with Tat Peptides
Wei Huang, Gabriele Varani and Gary P. Drobny
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja1051439/aop/images/medium/ja-2010-051439_0005.gif
Journal of the American Chemical Society
DOI: 10.1021/ja1051439
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[NMR paper] NMR study on the interaction between RPA and DNA decamer containing cis-syn cyclobuta
NMR study on the interaction between RPA and DNA decamer containing cis-syn cyclobutane pyrimidine dimer in the presence of XPA: implication for damage verification and strand-specific dual incision in nucleotide excision repair.
Related Articles NMR study on the interaction between RPA and DNA decamer containing cis-syn cyclobutane pyrimidine dimer in the presence of XPA: implication for damage verification and strand-specific dual incision in nucleotide excision repair.
Nucleic Acids Res. 2003 Aug 15;31(16):4747-54
Authors: Lee JH, Park CJ, Arunkumar...
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[NMR paper] NMR study of the interaction between the B domain of staphylococcal protein A and the
NMR study of the interaction between the B domain of staphylococcal protein A and the Fc portion of immunoglobulin G.
Related Articles NMR study of the interaction between the B domain of staphylococcal protein A and the Fc portion of immunoglobulin G.
Biochemistry. 1998 Jan 6;37(1):129-36
Authors: Gouda H, Shiraishi M, Takahashi H, Kato K, Torigoe H, Arata Y, Shimada I
The solution structure of the B domain of staphylococcal protein A (FB) complexed with the Fc fragment of immunoglobulin G (IgG) is reported. A previous NMR analysis has shown...
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[NMR paper] 19F NMR study of the interaction of fluoride ion with ribonucleotide reductase and me
19F NMR study of the interaction of fluoride ion with ribonucleotide reductase and methane monooxygenase.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles 19F NMR study of the interaction of fluoride ion with ribonucleotide reductase and methane monooxygenase.
Biochem Biophys Res Commun. 1993 Sep 15;195(2):594-9
Authors: Hamman S, Atta M, Ehrenberg A, Wilkins P, Dalton H, Béguin C, Fontecave M
The relaxation rates of fluoride, determined by 19F NMR spectroscopy, were...
Interaction study between HCV NS5A-D2 and NS5B using 19 F NMR
Linear Mode
