[NMR paper] Probing contacts of inhibitor locked in transition states in the catalytic triad of DENV2 type serine protease and its mutants by 1H, 19F and 15 N NMR spectroscopy.
Probing contacts of inhibitor locked in transition states in the catalytic triad of DENV2 type serine protease and its mutants by 1H, 19F and 15 N NMR spectroscopy.
Related ArticlesProbing contacts of inhibitor locked in transition states in the catalytic triad of DENV2 type serine protease and its mutants by 1H, 19F and 15 N NMR spectroscopy.
BMC Mol Cell Biol. 2020 May 25;21(1):38
Authors: Agback P, Woestenenk E, Agback T
Abstract
BACKGROUND: Detailed structural knowledge of enzyme-inhibitor complexes trapped in intermediate state is the key for a fundamental understanding of reaction mechanisms taking place in enzymes and is indispensable as a structure-guided drug design tool. Solution state NMR uniquely allows the study of active sites of enzymes in equilibrium between different tautomeric forms. In this study 1H, 19F and 15 N NMR spectroscopy has been used to probe the interaction contacts of inhibitors locked in transition states of the catalytic triad of a serine protease. It was demonstrated on the serotype II Dengue virus NS2B:NS3pro serine protease and its mutants, H51N and S135A, in complex with high-affinity ligands containing trifluoromethyl ketone (tfk) and boronic groups in the C-terminal of tetra-peptides.
RESULTS: Monitoring 19F resonances, shows that only one of the two isomers of the tfk tetra-peptide binds with NS2B:NS3pro and that access to the bulk of the active site is limited. Moreover, there were no bound water found in proximity of the active site for any of the ligands manifesting in a favorable condition for formation of low barrier hydrogen bonds (LBHB) in the catalytic triad. Based on this data we were able to identify a locked conformation of the protein active site. The data also indicates that the different parts of the binding site most likely act independently of each other.
CONCLUSIONS: Our reported findings increases the knowledge of the detailed function of the catalytic triad in serine proteases and could facilitate the development of rational structure based inhibitors that can selectively target the NS3 protease of Dengue type II (DENV2) virus. In addition the results shows the usefulness of probing active sites using 19F NMR spectroscopy.
[NMR paper] NMR resonance assignments of the catalytic domain of human serine/threonine phosphatase calcineurin in unligated and PVIVIT-peptide-bound states.
NMR resonance assignments of the catalytic domain of human serine/threonine phosphatase calcineurin in unligated and PVIVIT-peptide-bound states.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--production.springer.de-OnlineResources-Logos-springerlink.gif Related Articles NMR resonance assignments of the catalytic domain of human serine/threonine phosphatase calcineurin in unligated and PVIVIT-peptide-bound states.
Biomol NMR Assign. 2015 Apr;9(1):201-5
Authors: Takeuchi K, Sun ZY, Li S, Gal M, Wagner G
Abstract
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[NMR paper] Conformation of inhibitor-free HIV-1 protease derived from NMR spectroscopy in a weakly oriented solution.
Conformation of inhibitor-free HIV-1 protease derived from NMR spectroscopy in a weakly oriented solution.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--media.wiley.com-assets-2250-98-WileyOnlineLibrary_FullTextOnline_120x27.gif Related Articles Conformation of inhibitor-free HIV-1 protease derived from NMR spectroscopy in a weakly oriented solution.
Chembiochem. 2015 Jan 19;16(2):214-8
Authors: Roche J, Louis JM, Bax A
Abstract
Flexibility of the glycine-rich flaps is known to be essential for catalytic...
[NMR paper] Probing early misfolding events in prion protein mutants by NMR spectroscopy.
Probing early misfolding events in prion protein mutants by NMR spectroscopy.
Related Articles Probing early misfolding events in prion protein mutants by NMR spectroscopy.
Molecules. 2013;18(8):9451-76
Authors: Giachin G, Biljan I, Ilc G, Plavec J, Legname G
Abstract
The post-translational conversion of the ubiquitously expressed cellular form of the prion protein, PrPC, into its misfolded and pathogenic isoform, known as prion or PrPSc, plays a key role in prion diseases. These maladies are denoted transmissible spongiform...
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[NMR paper] Probing the cation binding modes of macrocyclic HCV protease inhibitor BILN 2061 by multinuclear NMR.
Probing the cation binding modes of macrocyclic HCV protease inhibitor BILN 2061 by multinuclear NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Probing the cation binding modes of macrocyclic HCV protease inhibitor BILN 2061 by multinuclear NMR.
J Pharm Biomed Anal. 2012 Nov;70:609-13
Authors: Busacca CA, Jones PJ, Campbell SJ, Saha AK, Gonnella NC, Senanayake CH
Abstract
The ability of the macrocyclic HCV protease inhibitor BILN 2061 to bind different...
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02-09-2013 12:18 AM
Probing water-protein contacts in a MMP-12/CGS27023A complex by nuclear magnetic resonance spectroscopy
Probing water-protein contacts in a MMP-12/CGS27023A complex by nuclear magnetic resonance spectroscopy
Abstract Using the case of the catalytic domain of MMP-12 in complex with the known inhibitor CGS27023A, a recently assembled 3D 15N-edited/14N,12C-filtered ROESY experiment is used to monitor and distinguish protein amide protons in fast exchange with bulk water from amide protons close to water molecules with longer residence times, the latter possibly reflecting water molecules of structural or functional importance. The 15N-edited/14N,12C-filtered ROESY spectra were compared to...
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Using the water signal to detect invisible exchanging protons in the catalytic triad of a serine protease
Using the water signal to detect invisible exchanging protons in the catalytic triad of a serine protease
Abstract Chemical Exchange Saturation Transfer (CEST) is an MRI approach that can indirectly detect exchange broadened protons that are invisible in traditional NMR spectra. We modified the CEST pulse sequence for use on high-resolution spectrometers and developed a quantitative approach for measuring exchange rates based upon CEST spectra. This new methodology was applied to the rapidly exchanging Hδ1 and Hε2 protons of His57 in the catalytic triad of bovine chymotrypsinogen-A...
Probing contacts of inhibitor locked in transition states in the catalytic triad of DENV2 type serine protease and its mutants by 1H, 19F and 15 N NMR spectroscopy.
Linear Mode
