5brb: Difference between revisions
New page: '''Unreleased structure''' The entry 5brb is ON HOLD Authors: Bandyopadhyay, D., Murthy, M.R.N., Balaram, H., Balaram, P. Description: Category: Unreleased Structures [[Category: ... |
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The | ==Crystal structure of Q64E mutant of Triosephosphate isomerase from Plasmodium falciparum== | ||
<StructureSection load='5brb' size='340' side='right'caption='[[5brb]], [[Resolution|resolution]] 2.53Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5brb]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Plasmodium_falciparum Plasmodium falciparum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5BRB OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5BRB FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.53Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NA:SODIUM+ION'>NA</scene></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=5brb FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5brb OCA], [https://pdbe.org/5brb PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5brb RCSB], [https://www.ebi.ac.uk/pdbsum/5brb PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5brb ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/TPIS_PLAFA TPIS_PLAFA] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Highly conserved residues in enzymes are often found to be clustered close to active sites, suggesting that functional constraints dictate the nature of amino acid residues accommodated at these sites. Using the Plasmodium falciparum Triosephosphate isomerase enzyme (PfTIM) as a template we have examined the effects of mutations at positions 64 and 75, which are not directly involved in the proton transfer cycle. Threonine (Thr) occurring at position 75 is completely conserved, while only Gln (Q) and Glu (E) are accommodated at position 64. Biophysical and kinetic data are reported for four T75 (T75S/V/C/N) and two Q64 (Q64N/E) mutants. The dimeric structure is weakened in the Q64E and Q64N mutants, whereas dimer integrity is unimpaired in all four T75 mutants. Measurement of the concentration dependence of enzyme activity permits an estimate of Kd values for dimer dissociation (Q64N = 73.7 +/- 9.2 nM and Q64E = 44.6 +/-8.4 nM). The T75S/V/C mutants have activities comparable to the wild type enzyme, while a four-fold drop is observed for T75N. All four T75 mutants show a dramatic fall in activity between 35 degrees - 45 degrees C. Crystal structure determination of the T75S/V/N mutants provides insights into the variations in local interactions, with the T75N mutant showing the largest changes. Hydrogen bond interactions determine dimer stability restricting the choice of residues at position 64 to Gln (Q) and Glu (E). At position 75, the overwhelming preference for Thr (T) may be dictated by the imperative of maintaining temperature stability of enzyme activity. This article is protected by copyright. All rights reserved. | |||
Probing the role of highly conserved residues in triosephosphate isomerase: analysis of site specific mutants at positions 64 and 75 in the Plasmodial enzyme.,Bandyopadhyay D, Murthy MR, Balaram H, Balaram P FEBS J. 2015 Jul 23. doi: 10.1111/febs.13384. PMID:26206206<ref>PMID:26206206</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: | <div class="pdbe-citations 5brb" style="background-color:#fffaf0;"></div> | ||
[[Category: Balaram | |||
[[Category: | ==See Also== | ||
[[Category: Murthy | *[[Triose phosphate isomerase 3D structures|Triose phosphate isomerase 3D structures]] | ||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Plasmodium falciparum]] | |||
[[Category: Balaram H]] | |||
[[Category: Balaram P]] | |||
[[Category: Bandyopadhyay D]] | |||
[[Category: Murthy MRN]] | |||
Latest revision as of 19:05, 8 November 2023
Crystal structure of Q64E mutant of Triosephosphate isomerase from Plasmodium falciparumCrystal structure of Q64E mutant of Triosephosphate isomerase from Plasmodium falciparum
Structural highlights
FunctionPublication Abstract from PubMedHighly conserved residues in enzymes are often found to be clustered close to active sites, suggesting that functional constraints dictate the nature of amino acid residues accommodated at these sites. Using the Plasmodium falciparum Triosephosphate isomerase enzyme (PfTIM) as a template we have examined the effects of mutations at positions 64 and 75, which are not directly involved in the proton transfer cycle. Threonine (Thr) occurring at position 75 is completely conserved, while only Gln (Q) and Glu (E) are accommodated at position 64. Biophysical and kinetic data are reported for four T75 (T75S/V/C/N) and two Q64 (Q64N/E) mutants. The dimeric structure is weakened in the Q64E and Q64N mutants, whereas dimer integrity is unimpaired in all four T75 mutants. Measurement of the concentration dependence of enzyme activity permits an estimate of Kd values for dimer dissociation (Q64N = 73.7 +/- 9.2 nM and Q64E = 44.6 +/-8.4 nM). The T75S/V/C mutants have activities comparable to the wild type enzyme, while a four-fold drop is observed for T75N. All four T75 mutants show a dramatic fall in activity between 35 degrees - 45 degrees C. Crystal structure determination of the T75S/V/N mutants provides insights into the variations in local interactions, with the T75N mutant showing the largest changes. Hydrogen bond interactions determine dimer stability restricting the choice of residues at position 64 to Gln (Q) and Glu (E). At position 75, the overwhelming preference for Thr (T) may be dictated by the imperative of maintaining temperature stability of enzyme activity. This article is protected by copyright. All rights reserved. Probing the role of highly conserved residues in triosephosphate isomerase: analysis of site specific mutants at positions 64 and 75 in the Plasmodial enzyme.,Bandyopadhyay D, Murthy MR, Balaram H, Balaram P FEBS J. 2015 Jul 23. doi: 10.1111/febs.13384. PMID:26206206[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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