8cek: Difference between revisions

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-'''Unreleased structure'''
-The entry 8cek is ON HOLD  until Paper Publication
+==Succinyl-CoA Reductase from Clostridium kluyveri (SucD) with NADPH==
+<StructureSection load='8cek' size='340' side='right'caption='[[8cek]], [[Resolution|resolution]] 2.15&Aring;' scene=''>
+== Structural highlights ==
+<table><tr><td colspan='2'>[[8cek]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Clostridium_kluyveri Clostridium kluyveri]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=8CEK OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=8CEK FirstGlance]. <br>
+</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</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=8cek FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=8cek OCA], [https://pdbe.org/8cek PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=8cek RCSB], [https://www.ebi.ac.uk/pdbsum/8cek PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=8cek ProSAT]</span></td></tr>
+</table>
+== Function ==
+[https://www.uniprot.org/uniprot/SUCD_CLOK5 SUCD_CLOK5] Catalyzes the reduction of succinate semialdehyde to succinyl-CoA. The enzyme is specific for succinate semialdehyde and succinyl-CoA, and only shows low activity with palmitoyl-CoA. There is no activity with NAD(+) as cosubstrate.
+<div style="background-color:#fffaf0;">
+== Publication Abstract from PubMed ==
+Succinyl-CoA reductase (SucD) is an acylating aldehyde reductase that catalyzes the NADPH-dependent reduction of succinyl-CoA to succinic semialdehyde. The reaction sequence from succinate to crotonyl-CoA is of particular interest for several new-to-nature CO(2)-fixation pathways, such as the crotonyl-CoA/ethylmalonyl-CoA/hydroxybutyryl-CoA (CETCH) cycle, in which SucD plays a key role. However, pathways like the CETCH cycle feature several CoA-ester intermediates, which could be potentially side substrates for this enzyme. Here, we show that the side reaction for most CETCH cycle metabolites is relatively small (&lt;2%) with the exception of mesaconyl-C1-CoA (16%), which represents a competing substrate in this pathway. We addressed this promiscuity by solving the crystal structure of a SucD of Clostridium kluyveri in complex with NADP(+) and mesaconyl-C1-CoA. We further identified two residues (Lys70 and Ser243) that coordinate mesaconyl-C1-CoA at the active site. We targeted those residues with site-directed mutagenesis to improve succinyl-CoA over mesaconyl-C1-CoA reduction. The best resulting SucD variant, K70R, showed a strongly reduced side activity for mesaconyl-C1-CoA, but the substitution also reduced the specific activity for succinyl-CoA by a factor of 10. Transferring the same mutations into a SucD homologue from Clostridium difficile similarly decreases the side reaction of this enzyme for mesaconyl-C1-CoA from 12 to 2%, notably without changing the catalytic efficiency for succinyl-CoA. Overall, our structure-based engineering efforts provided a highly specific enzyme of interest for several applications in biocatalysis and synthetic biology.
-Authors: Pfister, P., Diehl, C., Erb, T.J.
+Enhancing the Substrate Specificity of Clostridium Succinyl-CoA Reductase for Synthetic Biology and Biocatalysis.,Pfister P, Diehl C, Hammarlund E, Carrillo M, Erb TJ Biochemistry. 2023 May 19. doi: 10.1021/acs.biochem.3c00102. PMID:37207322<ref>PMID:37207322</ref>
-Description: Succinyl-CoA Reductase from Clostridium kluyveri (SucD) with NADPH
+From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
-[[Category: Unreleased Structures]]
+</div>
-[[Category: Pfister, P]]
+<div class="pdbe-citations 8cek" style="background-color:#fffaf0;"></div>
-[[Category: Erb, T.J]]
+== References ==
-[[Category: Diehl, C]]
+<references/>
+__TOC__
+</StructureSection>
+[[Category: Clostridium kluyveri]]
+[[Category: Large Structures]]
+[[Category: Diehl C]]
+[[Category: Erb TJ]]
+[[Category: Pfister P]]