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3vhq
From Proteopedia
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| - | '''Unreleased structure''' | ||
| - | + | ==Crystal structure of the Ca6 site mutant of Pro-SA-subtilisin== | |
| + | <StructureSection load='3vhq' size='340' side='right'caption='[[3vhq]], [[Resolution|resolution]] 2.15Å' scene=''> | ||
| + | == Structural highlights == | ||
| + | <table><tr><td colspan='2'>[[3vhq]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Pyrococcus_kodakaraensis_(strain_kod1) Pyrococcus kodakaraensis (strain kod1)]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3VHQ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3VHQ FirstGlance]. <br> | ||
| + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene></td></tr> | ||
| + | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[2e1p|2e1p]], [[2zwo|2zwo]], [[2zwp|2zwp]]</div></td></tr> | ||
| + | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">TK1675 ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=69014 Pyrococcus kodakaraensis (strain KOD1)])</td></tr> | ||
| + | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[https://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [https://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] </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=3vhq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3vhq OCA], [https://pdbe.org/3vhq PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3vhq RCSB], [https://www.ebi.ac.uk/pdbsum/3vhq PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3vhq ProSAT]</span></td></tr> | ||
| + | </table> | ||
| + | == Function == | ||
| + | [[https://www.uniprot.org/uniprot/TKSU_THEKO TKSU_THEKO]] Has a broad substrate specificity with a slight preference to large hydrophobic amino acid residues at the P1 position. | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Tk-subtilisin, a hyperthermostable subtilisin-like serine protease from Thermococcus kodakarensis, matures from the inactive precursor, Pro-Tk-subtilisin (Pro-TKS), upon autoprocessing and degradation of the propeptide (Tkpro). It contains seven Ca(2+) ions. Four of them (Ca2-Ca5) are responsible for folding of Tk-subtilisin. In this study, to clarify the role of the other three Ca(2+) ions (Ca1, Ca6, and Ca7), we constructed Pro-TKS derivatives lacking the Ca1 ion (Pro-TKS/DeltaCa1), Ca6 ion (Pro-TKS/DeltaCa6), and Ca7 ion (Pro-TKS/DeltaCa7), and their active site mutants (Pro-S324A/DeltaCa1, Pro-S324A/DeltaCa6, and Pro-S324A/DeltaCa7, respectively). Pro-TKS/DeltaCa6 and Pro-TKS/DeltaCa7 fully matured into their active forms upon incubation at 80 degrees C for 30 min as did Pro-TKS. The mature enzymes were as active as Tk-subtilisin at 80 degrees C, indicating that the Ca6 and Ca7 ions are not important for activity. In contrast, Pro-TKS/DeltaCa1 matured poorly at 80 degrees C because of the instability of its mature domain. The enzymatic activity of Tk-subtilisin/DeltaCa1 was determined to be 50% of that of Tk-subtilisin using the refolded protein. This result suggests that the Ca1 ion is required for the maximal activity of Tk-subtilisin. The refolding rates of all Pro-S324A derivatives were comparable to that of Pro-S324A (active site mutant of Pro-TKS), indicating that these Ca(2+) ions are not needed for folding of Tk-subtilisin. The stabilities of Pro-S324A/DeltaCa1 and Pro-S324A/DeltaCa6 were decreased by 26.6 and 11.7 degrees C, respectively, in T(m) compared to that of Pro-S324A. The half-lives of Tk-subtilisin/DeltaCa6 and Tk-subtilisin/DeltaCa7 at 95 degrees C were 8- and 4-fold lower than that of Tk-subtilisin, respectively. These results suggest that the Ca1, Ca6, and Ca7 ions, especially the Ca1 ion, contribute to the hyperthermostabilization of Tk-subtilisin. | ||
| - | + | Requirement of Ca(2+) Ions for the Hyperthermostability of Tk-Subtilisin from Thermococcus kodakarensis.,Uehara R, Takeuchi Y, Tanaka SI, Takano K, Koga Y, Kanaya S Biochemistry. 2012 Jun 19. PMID:22686281<ref>PMID:22686281</ref> | |
| - | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |
| + | </div> | ||
| + | <div class="pdbe-citations 3vhq" style="background-color:#fffaf0;"></div> | ||
| + | |||
| + | ==See Also== | ||
| + | *[[Subtilisin 3D structures|Subtilisin 3D structures]] | ||
| + | == References == | ||
| + | <references/> | ||
| + | __TOC__ | ||
| + | </StructureSection> | ||
| + | [[Category: Large Structures]] | ||
| + | [[Category: Subtilisin]] | ||
| + | [[Category: Kanaya, S]] | ||
| + | [[Category: Koga, Y]] | ||
| + | [[Category: Matsumura, H]] | ||
| + | [[Category: Takano, K]] | ||
| + | [[Category: Takeuchi, Y]] | ||
| + | [[Category: Tanaka, S]] | ||
| + | [[Category: Uehara, R]] | ||
| + | [[Category: Hydrolase]] | ||
| + | [[Category: Proteolysis]] | ||
Current revision
Crystal structure of the Ca6 site mutant of Pro-SA-subtilisin
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Categories: Large Structures | Subtilisin | Kanaya, S | Koga, Y | Matsumura, H | Takano, K | Takeuchi, Y | Tanaka, S | Uehara, R | Hydrolase | Proteolysis
