1zvk

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(New page: 200px<br /><applet load="1zvk" size="350" color="white" frame="true" align="right" spinBox="true" caption="1zvk, resolution 2.04&Aring;" /> '''Structure of Double ...)
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[[Image:1zvk.gif|left|200px]]<br /><applet load="1zvk" size="350" color="white" frame="true" align="right" spinBox="true"
 
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caption="1zvk, resolution 2.04&Aring;" />
 
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'''Structure of Double mutant, D164N, E78H of Kumamolisin-As'''<br />
 
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==Overview==
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==Structure of Double mutant, D164N, E78H of Kumamolisin-As==
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Kumamolisin-As is an acid collagenase with a subtilisin-like fold. Its, active site contains a unique catalytic triad, Ser278-Glu78-Asp82, and a, putative transition-state stabilizing residue, Asp164. In this study, the, mutants D164N and E78H/D164N were engineered in order to replace parts of, the catalytic machinery of kumamolisin-As with the residues found in the, equivalent positions in subtilisin. Unlike the wild-type and D164N, proenzymes, which undergo instantaneous processing to produce their 37-kDa, mature forms, the expressed E78H/D164N proenzyme exists as an equilibrated, mixture of the nicked and intact forms of the precursor. X-ray, crystallographic structures of the mature forms of the two mutants showed, that, in each of them, the catalytic Ser278 makes direct hydrogen bonds, with the side chain of Asn164. In addition, His78 of the double mutant is, distant from Ser278 and Asp82, and the catalytic triad no longer exists., Consistent with these structural alterations around the active site, these, mutants showed only low catalytic activity (relative k(cat) at pH 4.0 1.3%, for D164N and 0.0001% for E78H/D164N). pH-dependent kinetic studies showed, that the single D164N substitution did not significantly alter the, logk(cat) vs. pH and log(k(cat)/Km) vs. pH profiles of the enzyme. In, contrast, the double mutation resulted in a dramatic switch of the, logk(cat) vs. pH profile to one that was consistent with catalysis by, means of the Ser278-His78 dyad and Asn164, which may also account for the, observed ligation/cleavage equilibrium of the precursor of E78H/D164N., These results corroborate the mechanistic importance of the, glutamate-mediated catalytic triad and oxyanion-stabilizing aspartic acid, residue for low-pH peptidase activity of the enzyme.
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<StructureSection load='1zvk' size='340' side='right'caption='[[1zvk]], [[Resolution|resolution]] 2.04&Aring;' scene=''>
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== Structural highlights ==
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==About this Structure==
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<table><tr><td colspan='2'>[[1zvk]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Alicyclobacillus_sendaiensis Alicyclobacillus sendaiensis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1ZVK OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1ZVK FirstGlance]. <br>
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1ZVK is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Alicyclobacillus_sendaiensis Alicyclobacillus sendaiensis] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1ZVK OCA].
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.04&#8491;</td></tr>
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<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>
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==Reference==
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1zvk FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1zvk OCA], [https://pdbe.org/1zvk PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1zvk RCSB], [https://www.ebi.ac.uk/pdbsum/1zvk PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1zvk ProSAT]</span></td></tr>
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Processing, catalytic activity and crystal structures of kumamolisin-As with an engineered active site., Okubo A, Li M, Ashida M, Oyama H, Gustchina A, Oda K, Dunn BM, Wlodawer A, Nakayama T, FEBS J. 2006 Jun;273(11):2563-76. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=16704427 16704427]
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/Q8GB88_9BACL Q8GB88_9BACL]
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/zv/1zvk_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1zvk ConSurf].
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<div style="clear:both"></div>
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__TOC__
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</StructureSection>
[[Category: Alicyclobacillus sendaiensis]]
[[Category: Alicyclobacillus sendaiensis]]
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[[Category: Single protein]]
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[[Category: Large Structures]]
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[[Category: Gustchina, A.]]
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[[Category: Gustchina A]]
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[[Category: Li, M.]]
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[[Category: Li M]]
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[[Category: Nakayama, T.]]
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[[Category: Nakayama T]]
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[[Category: Wlodawer, A.]]
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[[Category: Wlodawer A]]
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[[Category: CA]]
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[[Category: d164n]]
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[[Category: e78h]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jan 29 17:43:47 2008''
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Current revision

Structure of Double mutant, D164N, E78H of Kumamolisin-As

PDB ID 1zvk

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