1n2t

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(New page: 200px<br /><applet load="1n2t" size="450" color="white" frame="true" align="right" spinBox="true" caption="1n2t, resolution 2.00&Aring;" /> '''C-DES Mutant K223A w...)
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[[Image:1n2t.jpg|left|200px]]<br /><applet load="1n2t" size="450" color="white" frame="true" align="right" spinBox="true"
 
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caption="1n2t, resolution 2.00&Aring;" />
 
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'''C-DES Mutant K223A with GLY Covalenty Linked to the PLP-cofactor'''<br />
 
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==Overview==
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==C-DES Mutant K223A with GLY Covalenty Linked to the PLP-cofactor==
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The cystine lyase (C-DES) of Synechocystis is a, pyridoxal-5'-phosphate-dependent enzyme distantly related to the family of, NifS-like proteins. The crystal structure of an N-terminal modified, variant has recently been determined. Herein, the reactivity of this, enzyme variant was investigated spectroscopically in solution and in the, crystalline state to follow the course of the reaction and to determine, the catalytic mechanism on a molecular level. Using the stopped-flow, technique, the reaction with the preferred substrate cystine was found to, follow biphasic kinetics leading to the formation of absorbing species at, 338 and 470 nm, attributed to the external aldimine and the, alpha-aminoacrylate; the reaction with cysteine also exhibited biphasic, behavior but only the external aldimine accumulated. The same reaction, intermediates were formed in crystals as seen by polarized absorption, microspectrophotometry, thus indicating that C-DES is catalytically, competent in the crystalline state. The three-dimensional structure of the, catalytically inactive mutant C-DES(K223A) in the presence of cystine, showed the formation of an external aldimine species, in which two, alternate conformations of the substrate were observed. The combined, results allow a catalytic mechanism to be proposed involving interactions, between cystine and the active site residues Arg-360, Arg-369, and, Trp-251*; these residues reorient during the beta-elimination reaction, leading to the formation of a hydrophobic pocket that stabilizes the, enolimine tautomer of the aminoacrylate and the cysteine persulfide, product.
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<StructureSection load='1n2t' size='340' side='right'caption='[[1n2t]], [[Resolution|resolution]] 2.00&Aring;' scene=''>
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== Structural highlights ==
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==About this Structure==
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<table><tr><td colspan='2'>[[1n2t]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Synechocystis_sp._PCC_6714 Synechocystis sp. PCC 6714]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1N2T OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1N2T FirstGlance]. <br>
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1N2T is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Synechocystis_sp._pcc_6803 Synechocystis sp. pcc 6803] with K, PLP and GLY as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1N2T 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&#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=GLY:GLYCINE'>GLY</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PLP:PYRIDOXAL-5-PHOSPHATE'>PLP</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=1n2t FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1n2t OCA], [https://pdbe.org/1n2t PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1n2t RCSB], [https://www.ebi.ac.uk/pdbsum/1n2t PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1n2t ProSAT]</span></td></tr>
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Snapshots of the cystine lyase C-DES during catalysis. Studies in solution and in the crystalline state., Kaiser JT, Bruno S, Clausen T, Huber R, Schiaretti F, Mozzarelli A, Kessler D, J Biol Chem. 2003 Jan 3;278(1):357-65. Epub 2002 Oct 16. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=12386155 12386155]
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</table>
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[[Category: Single protein]]
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== Function ==
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[[Category: Synechocystis sp. pcc 6803]]
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[https://www.uniprot.org/uniprot/Q9ZHG9_SYNY4 Q9ZHG9_SYNY4]
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[[Category: Bruno, S.]]
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== Evolutionary Conservation ==
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[[Category: Clausen, T.]]
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[[Image:Consurf_key_small.gif|200px|right]]
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[[Category: Huber, R.]]
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Check<jmol>
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[[Category: Kaiser, J.T.]]
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<jmolCheckbox>
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[[Category: Kessler, D.]]
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/n2/1n2t_consurf.spt"</scriptWhenChecked>
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[[Category: Mozzarelli, A.]]
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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[[Category: Schiaretti, F.]]
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<text>to colour the structure by Evolutionary Conservation</text>
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[[Category: GLY]]
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</jmolCheckbox>
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[[Category: K]]
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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=1n2t ConSurf].
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[[Category: PLP]]
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<div style="clear:both"></div>
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[[Category: fes cluster biosynthesis]]
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__TOC__
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[[Category: inactive mutant]]
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</StructureSection>
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[[Category: nifs]]
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[[Category: Large Structures]]
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[[Category: pyridoxal 5'-phosphate]]
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[[Category: Synechocystis sp. PCC 6714]]
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[[Category: Bruno S]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Sat Nov 24 22:27:56 2007''
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[[Category: Clausen T]]
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[[Category: Huber R]]
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[[Category: Kaiser JT]]
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[[Category: Kessler D]]
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[[Category: Mozzarelli A]]
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[[Category: Schiaretti F]]

Current revision

C-DES Mutant K223A with GLY Covalenty Linked to the PLP-cofactor

PDB ID 1n2t

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