6pcc: Difference between revisions
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==Crystal structure of beta-ketoadipyl-CoA thiolase mutant (H356A) in complex hexanoyl coenzyme A== | ==Crystal structure of beta-ketoadipyl-CoA thiolase mutant (H356A) in complex hexanoyl coenzyme A== | ||
<StructureSection load='6pcc' size='340' side='right'caption='[[6pcc]]' scene=''> | <StructureSection load='6pcc' size='340' side='right'caption='[[6pcc]], [[Resolution|resolution]] 1.96Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6PCC OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=6PCC FirstGlance]. <br> | <table><tr><td colspan='2'>[[6pcc]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Psepk Psepk]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6PCC OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=6PCC FirstGlance]. <br> | ||
</td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=6pcc FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6pcc OCA], [http://pdbe.org/6pcc PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6pcc RCSB], [http://www.ebi.ac.uk/pdbsum/6pcc PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6pcc ProSAT]</span></td></tr> | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=COA:COENZYME+A'>COA</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=O8Y:hexanal'>O8Y</scene></td></tr> | ||
<tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CSO:S-HYDROXYCYSTEINE'>CSO</scene></td></tr> | |||
<tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">pcaF-I, PP_1377 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=160488 PSEPK])</td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=6pcc FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6pcc OCA], [http://pdbe.org/6pcc PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6pcc RCSB], [http://www.ebi.ac.uk/pdbsum/6pcc PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6pcc ProSAT]</span></td></tr> | |||
</table> | </table> | ||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Thiolases are a well characterized family of enzymes with two distinct categories: degradative, beta-ketoadipyl-CoA thiolases and biosynthetic, acetoacetyl-CoA thiolases. Both classes share an identical catalytic triad but catalyze reactions in opposite directions. Moreover, it is established that in contrast to the biosynthetic thiolases the degradative thiolases can accept substrates with broad chain lengths. Hitherto, no residue or structural pattern has been recognized that might help to discern the two thiolases, here we exploit, a tetrameric degradative thiolase from Pseudomonas putida KT2440 annotated as PcaF, as a model system to understand features which distinguishes the two classes using structural studies and bioinformatics analyses. Degradative thiolases have different active site architecture when compared to biosynthetic thiolases, demonstrating the dissimilar chemical nature of the active site architecture. Both thiolases deploy different "anchoring residues" to tether the large Coenzyme A (CoA) or CoA derivatives. Interestingly, the H356 of the catalytic triad in PcaF is directly involved in tethering the CoA/CoA derivatives into the active site and we were able to trap a gridlocked thiolase structure of the H356A mutant, where the CoA was found to be covalently linked to the catalytic cysteine residue, inhibiting the overall reaction. Further, X-ray structures with two long chain CoA derivatives, hexanal-CoA and octanal-CoA helped in delineating the long tunnel of 235 A(2) surface area in PcaF and led to identification of a unique covering loop exclusive to degradative thiolases that plays an active role in determining the tunnel length and the nature of the binding substrate. | |||
Structural basis for differentiation between two classes of thiolase: Degradative vs biosynthetic thiolase.,Bhaskar S, Steer DL, Anand R, Panjikar S J Struct Biol X. 2020 Jan 3;4:100018. doi: 10.1016/j.yjsbx.2019.100018., eCollection 2020. PMID:32647822<ref>PMID:32647822</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 6pcc" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Panjikar S]] | [[Category: Psepk]] | ||
[[Category: Sukritee B]] | [[Category: Panjikar, S]] | ||
[[Category: Sukritee, B]] | |||
[[Category: Aromatic pollutant catabolism]] | |||
[[Category: Degradative enzyme]] | |||
[[Category: Thiolase]] | |||
[[Category: Transferase]] | |||
Revision as of 14:23, 22 July 2020
Crystal structure of beta-ketoadipyl-CoA thiolase mutant (H356A) in complex hexanoyl coenzyme ACrystal structure of beta-ketoadipyl-CoA thiolase mutant (H356A) in complex hexanoyl coenzyme A
Structural highlights
Publication Abstract from PubMedThiolases are a well characterized family of enzymes with two distinct categories: degradative, beta-ketoadipyl-CoA thiolases and biosynthetic, acetoacetyl-CoA thiolases. Both classes share an identical catalytic triad but catalyze reactions in opposite directions. Moreover, it is established that in contrast to the biosynthetic thiolases the degradative thiolases can accept substrates with broad chain lengths. Hitherto, no residue or structural pattern has been recognized that might help to discern the two thiolases, here we exploit, a tetrameric degradative thiolase from Pseudomonas putida KT2440 annotated as PcaF, as a model system to understand features which distinguishes the two classes using structural studies and bioinformatics analyses. Degradative thiolases have different active site architecture when compared to biosynthetic thiolases, demonstrating the dissimilar chemical nature of the active site architecture. Both thiolases deploy different "anchoring residues" to tether the large Coenzyme A (CoA) or CoA derivatives. Interestingly, the H356 of the catalytic triad in PcaF is directly involved in tethering the CoA/CoA derivatives into the active site and we were able to trap a gridlocked thiolase structure of the H356A mutant, where the CoA was found to be covalently linked to the catalytic cysteine residue, inhibiting the overall reaction. Further, X-ray structures with two long chain CoA derivatives, hexanal-CoA and octanal-CoA helped in delineating the long tunnel of 235 A(2) surface area in PcaF and led to identification of a unique covering loop exclusive to degradative thiolases that plays an active role in determining the tunnel length and the nature of the binding substrate. Structural basis for differentiation between two classes of thiolase: Degradative vs biosynthetic thiolase.,Bhaskar S, Steer DL, Anand R, Panjikar S J Struct Biol X. 2020 Jan 3;4:100018. doi: 10.1016/j.yjsbx.2019.100018., eCollection 2020. PMID:32647822[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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