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| | ==Biochemical and structural characterization of mycobacterial aspartyl-tRNA synthetase AspS, a promising TB drug target== | | ==Biochemical and structural characterization of mycobacterial aspartyl-tRNA synthetase AspS, a promising TB drug target== |
| - | <StructureSection load='4rmf' size='340' side='right' caption='[[4rmf]], [[Resolution|resolution]] 2.40Å' scene=''> | + | <StructureSection load='4rmf' size='340' side='right'caption='[[4rmf]], [[Resolution|resolution]] 2.40Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4rmf]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4RMF OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4RMF FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4rmf]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Mycolicibacterium_smegmatis_MC2_155 Mycolicibacterium smegmatis MC2 155]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4RMF OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4RMF FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=3SY:2,2-BIS(HYDROXYMETHYL)PROPANE-1,3-DIOL'>3SY</scene>, <scene name='pdbligand=FMT:FORMIC+ACID'>FMT</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.4Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Aspartate--tRNA(Asn)_ligase Aspartate--tRNA(Asn) ligase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=6.1.1.23 6.1.1.23] </span></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=3SY:2,2-BIS(HYDROXYMETHYL)PROPANE-1,3-DIOL'>3SY</scene>, <scene name='pdbligand=FMT:FORMIC+ACID'>FMT</scene></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4rmf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4rmf OCA], [http://pdbe.org/4rmf PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4rmf RCSB], [http://www.ebi.ac.uk/pdbsum/4rmf PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4rmf ProSAT]</span></td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=4rmf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4rmf OCA], [https://pdbe.org/4rmf PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4rmf RCSB], [https://www.ebi.ac.uk/pdbsum/4rmf PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4rmf ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/SYDND_MYCS2 SYDND_MYCS2]] Aspartyl-tRNA synthetase with relaxed tRNA specificity since it is able to aspartylate not only its cognate tRNA(Asp) but also tRNA(Asn). Reaction proceeds in two steps: aspartate is first activated by ATP to form Asp-AMP and then transferred to the acceptor end of tRNA(Asp/Asn).[HAMAP-Rule:MF_00044] | + | [https://www.uniprot.org/uniprot/SYDND_MYCS2 SYDND_MYCS2] Aspartyl-tRNA synthetase with relaxed tRNA specificity since it is able to aspartylate not only its cognate tRNA(Asp) but also tRNA(Asn). Reaction proceeds in two steps: aspartate is first activated by ATP to form Asp-AMP and then transferred to the acceptor end of tRNA(Asp/Asn).[HAMAP-Rule:MF_00044] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </div> | | </div> |
| | <div class="pdbe-citations 4rmf" style="background-color:#fffaf0;"></div> | | <div class="pdbe-citations 4rmf" style="background-color:#fffaf0;"></div> |
| | + | |
| | + | ==See Also== |
| | + | *[[Aminoacyl tRNA synthetase 3D structures|Aminoacyl tRNA synthetase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Besra, G S]] | + | [[Category: Large Structures]] |
| - | [[Category: Cox, J A.G]] | + | [[Category: Mycolicibacterium smegmatis MC2 155]] |
| - | [[Category: Futterer, K]] | + | [[Category: Besra GS]] |
| - | [[Category: Gurcha, S S]] | + | [[Category: Cox JAG]] |
| - | [[Category: Veeraraghavan, U]] | + | [[Category: Futterer K]] |
| - | [[Category: Alpha and beta protein]] | + | [[Category: Gurcha SS]] |
| - | [[Category: Ligase]] | + | [[Category: Veeraraghavan U]] |
| - | [[Category: Trna]]
| + | |
| - | [[Category: Trna synthetase]]
| + | |
| Structural highlights
Function
SYDND_MYCS2 Aspartyl-tRNA synthetase with relaxed tRNA specificity since it is able to aspartylate not only its cognate tRNA(Asp) but also tRNA(Asn). Reaction proceeds in two steps: aspartate is first activated by ATP to form Asp-AMP and then transferred to the acceptor end of tRNA(Asp/Asn).[HAMAP-Rule:MF_00044]
Publication Abstract from PubMed
The human pathogen Mycobacterium tuberculosis is the causative agent of pulmonary tuberculosis (TB), a disease with high worldwide mortality rates. Current treatment programs are under significant threat from multi-drug and extensively-drug resistant strains of M. tuberculosis, and it is essential to identify new inhibitors and their targets. We generated spontaneous resistant mutants in Mycobacterium bovis BCG in the presence of 10x the minimum inhibitory concentration (MIC) of compound 1, a previously identified potent inhibitor of mycobacterial growth in culture. Whole genome sequencing of two resistant mutants revealed in one case a single nucleotide polymorphism in the gene aspS at 535GAC>535AAC (D179N), while in the second mutant a single nucleotide polymorphism was identified upstream of the aspS promoter region. We probed whole cell target engagement by overexpressing either M. bovis BCG aspS or Mycobacterium smegmatis aspS, which resulted in a ten-fold and greater than ten-fold increase, respectively, of the MIC against compound 1. To analyse the impact of inhibitor 1 on M. tuberculosis AspS (Mt-AspS) activity we over-expressed, purified and characterised the kinetics of this enzyme using a robust tRNA-independent assay adapted to a high-throughput screening format. Finally, to aid hit-to-lead optimization, the crystal structure of apo M. smegmatis AspS was determined to a resolution of 2.4 A.
Biochemical and Structural Characterization of Mycobacterial Aspartyl-tRNA Synthetase AspS, a Promising TB Drug Target.,Gurcha SS, Usha V, Cox JA, Futterer K, Abrahams KA, Bhatt A, Alderwick LJ, Reynolds RC, Loman NJ, Nataraj V, Alemparte C, Barros D, Lloyd AJ, Ballell L, Hobrath JV, Besra GS PLoS One. 2014 Nov 19;9(11):e113568. doi: 10.1371/journal.pone.0113568., eCollection 2014. PMID:25409504[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Gurcha SS, Usha V, Cox JA, Futterer K, Abrahams KA, Bhatt A, Alderwick LJ, Reynolds RC, Loman NJ, Nataraj V, Alemparte C, Barros D, Lloyd AJ, Ballell L, Hobrath JV, Besra GS. Biochemical and Structural Characterization of Mycobacterial Aspartyl-tRNA Synthetase AspS, a Promising TB Drug Target. PLoS One. 2014 Nov 19;9(11):e113568. doi: 10.1371/journal.pone.0113568., eCollection 2014. PMID:25409504 doi:http://dx.doi.org/10.1371/journal.pone.0113568
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