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| | ==Crystal structure of the Ca6 site mutant of Pro-SA-subtilisin== | | ==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=''> | + | <StructureSection load='3vhq' size='340' side='right'caption='[[3vhq]], [[Resolution|resolution]] 2.15Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3vhq]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Thermococcus_kodakarensis Thermococcus kodakarensis]. 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 [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3VHQ FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3vhq]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Thermococcus_kodakarensis_KOD1 Thermococcus kodakarensis 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><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CA:CALCIUM+ION'>CA</scene><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]] 2.15Å</td></tr> |
| - | <tr><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2e1p|2e1p]], [[2zwo|2zwo]], [[2zwp|2zwp]]</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><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">TK1675 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=311400 Thermococcus kodakarensis])</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> |
| - | <tr><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] </span></td></tr>
| + | </table> |
| - | <tr><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3vhq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3vhq OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3vhq RCSB], [http://www.ebi.ac.uk/pdbsum/3vhq PDBsum]</span></td></tr> | + | == Function == |
| - | <table> | + | [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;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | </div> | | </div> |
| | + | <div class="pdbe-citations 3vhq" style="background-color:#fffaf0;"></div> |
| | | | |
| | ==See Also== | | ==See Also== |
| - | *[[Subtilisin|Subtilisin]] | + | *[[Subtilisin 3D structures|Subtilisin 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Subtilisin]] | + | [[Category: Large Structures]] |
| - | [[Category: Thermococcus kodakarensis]] | + | [[Category: Thermococcus kodakarensis KOD1]] |
| - | [[Category: Kanaya, S.]] | + | [[Category: Kanaya S]] |
| - | [[Category: Koga, Y.]] | + | [[Category: Koga Y]] |
| - | [[Category: Matsumura, H.]] | + | [[Category: Matsumura H]] |
| - | [[Category: Takano, K.]] | + | [[Category: Takano K]] |
| - | [[Category: Takeuchi, Y.]] | + | [[Category: Takeuchi Y]] |
| - | [[Category: Tanaka, S.]] | + | [[Category: Tanaka S]] |
| - | [[Category: Uehara, R.]] | + | [[Category: Uehara R]] |
| - | [[Category: Hydrolase]]
| + | |
| - | [[Category: Proteolysis]]
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| Structural highlights
Function
TKSU_THEKO Has a broad substrate specificity with a slight preference to large hydrophobic amino acid residues at the P1 position.
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[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Uehara R, Takeuchi Y, Tanaka SI, Takano K, Koga Y, Kanaya S. Requirement of Ca(2+) Ions for the Hyperthermostability of Tk-Subtilisin from Thermococcus kodakarensis. Biochemistry. 2012 Jun 19. PMID:22686281 doi:10.1021/bi300427u
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