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4pcf
From Proteopedia
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==Structure-based protein engineering of a monomeric triosephosphate isomerase towards changing substrate specificity== | ==Structure-based protein engineering of a monomeric triosephosphate isomerase towards changing substrate specificity== | ||
<StructureSection load='4pcf' size='340' side='right' caption='[[4pcf]], [[Resolution|resolution]] 2.71Å' scene=''> | <StructureSection load='4pcf' size='340' side='right' caption='[[4pcf]], [[Resolution|resolution]] 2.71Å' scene=''> | ||
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</td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5tim|5tim]], [[1tri|1tri]], [[2vek|2vek]], [[2vel|2vel]], [[1ag1|1ag1]], [[1dkw|1dkw]], [[1iig|1iig]], [[1ml1|1ml1]], [[1mss|1mss]], [[2v0t|2v0t]], [[2v5l|2v5l]], [[6tim|6tim]], [[2vem|2vem]], [[2ven|2ven]], [[2vei|2vei]], [[3tim|3tim]]</td></tr> | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[5tim|5tim]], [[1tri|1tri]], [[2vek|2vek]], [[2vel|2vel]], [[1ag1|1ag1]], [[1dkw|1dkw]], [[1iig|1iig]], [[1ml1|1ml1]], [[1mss|1mss]], [[2v0t|2v0t]], [[2v5l|2v5l]], [[6tim|6tim]], [[2vem|2vem]], [[2ven|2ven]], [[2vei|2vei]], [[3tim|3tim]]</td></tr> | ||
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Triose-phosphate_isomerase Triose-phosphate isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.1.1 5.3.1.1] </span></td></tr> | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Triose-phosphate_isomerase Triose-phosphate isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.3.1.1 5.3.1.1] </span></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=4pcf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4pcf OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4pcf RCSB], [http://www.ebi.ac.uk/pdbsum/4pcf PDBsum]</span></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=4pcf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4pcf OCA], [http://pdbe.org/4pcf PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4pcf RCSB], [http://www.ebi.ac.uk/pdbsum/4pcf PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4pcf ProSAT]</span></td></tr> |
</table> | </table> | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | The crystal structures are described of two variants of A-TIM: Ma18 (2.7 A resolution) and Ma21 (1.55 A resolution). A-TIM is a monomeric loop-deletion variant of triosephosphate isomerase (TIM) which has lost the TIM catalytic properties. Ma18 and Ma21 were identified after extensive directed-evolution selection experiments using an Escherichia coli L-arabinose isomerase knockout strain expressing a randomly mutated A-TIM gene. These variants facilitate better growth of the Escherichia coli selection strain in medium supplemented with 40 mM L-arabinose. Ma18 and Ma21 differ from A-TIM by four and one point mutations, respectively. Ma18 and Ma21 are more stable proteins than A-TIM, as judged from CD melting experiments. Like A-TIM, both proteins are monomeric in solution. In the Ma18 crystal structure loop 6 is open and in the Ma21 crystal structure loop 6 is closed, being stabilized by a bound glycolate molecule. The crystal structures show only small differences in the active site compared with A-TIM. In the case of Ma21 it is observed that the point mutation (Q65L) contributes to small structural rearrangements near Asn11 of loop 1, which correlate with different ligand-binding properties such as a loss of citrate binding in the active site. The Ma21 structure also shows that its Leu65 side chain is involved in van der Waals interactions with neighbouring hydrophobic side-chain moieties, correlating with its increased stability. The experimental data suggest that the increased stability and solubility properties of Ma21 and Ma18 compared with A-TIM cause better growth of the selection strain when coexpressing Ma21 and Ma18 instead of A-TIM. | ||
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| + | Crystal structures of two monomeric triosephosphate isomerase variants identified via a directed-evolution protocol selecting for L-arabinose isomerase activity.,Krause M, Kiema TR, Neubauer P, Wierenga RK Acta Crystallogr F Struct Biol Commun. 2016 Jun 1;72(Pt 6):490-9. doi:, 10.1107/S2053230X16007548. Epub 2016 May 23. PMID:27303904<ref>PMID:27303904</ref> | ||
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| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
| + | </div> | ||
| + | <div class="pdbe-citations 4pcf" style="background-color:#fffaf0;"></div> | ||
| + | == References == | ||
| + | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
Revision as of 17:47, 12 July 2016
Structure-based protein engineering of a monomeric triosephosphate isomerase towards changing substrate specificity
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