1ojj

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[[Image:1ojj.gif|left|200px]]<br />
 
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<applet load="1ojj" size="450" color="white" frame="true" align="right" spinBox="true"
 
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caption="1ojj, resolution 1.40&Aring;" />
 
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'''ANATOMY OF GLYCOSYNTHESIS: STRUCTURE AND KINETICS OF THE HUMICOLA INSOLENS CEL7BE197A AND E197S GLYCOSYNTHASE MUTANTS'''<br />
 
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==Overview==
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==Anatomy of glycosynthesis: Structure and kinetics of the Humicola insolens Cel7BE197A and E197S glycosynthase mutants==
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The formation of glycoconjugates and oligosaccharides remains one of the, most challenging chemical syntheses. Chemo-enzymatic routes using, retaining glycosidases have been successfully harnessed but require tight, kinetic or thermodynamic control. "Glycosynthases," specifically, engineered glycosidases that catalyze the formation of glycosidic bonds, from glycosyl donor and acceptor alcohol, are an emerging range of, synthetic tools in which catalytic nucleophile mutants are harnessed, together with glycosyl fluoride donors to generate powerful and versatile, catalysts. Here we present the structural and kinetic dissection of the, Humicola insolens Cel7B glycosynthases in which the nucleophile of the, wild-type enzyme is mutated to alanine and serine (E197A and E197S). 3-D, structures reveal the acceptor and donor subsites and the basis for, substrate inhibition. Kinetic analysis shows that the E197S mutant is, considerably more active than the corresponding alanine mutant due to a, 40-fold increase in k(cat).
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<StructureSection load='1ojj' size='340' side='right'caption='[[1ojj]], [[Resolution|resolution]] 1.40&Aring;' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[1ojj]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Humicola_insolens Humicola insolens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1OJJ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1OJJ FirstGlance]. <br>
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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]] 1.4&#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=BGC:BETA-D-GLUCOSE'>BGC</scene>, <scene name='pdbligand=GAL:BETA-D-GALACTOSE'>GAL</scene>, <scene name='pdbligand=GLC:ALPHA-D-GLUCOSE'>GLC</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=PCA:PYROGLUTAMIC+ACID'>PCA</scene>, <scene name='pdbligand=PRD_900004:beta-lactose'>PRD_900004</scene>, <scene name='pdbligand=PRD_900008:alpha-lactose'>PRD_900008</scene></td></tr>
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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=1ojj FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1ojj OCA], [https://pdbe.org/1ojj PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1ojj RCSB], [https://www.ebi.ac.uk/pdbsum/1ojj PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1ojj ProSAT]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/GUN1_HUMIN GUN1_HUMIN] The biological conversion of cellulose to glucose generally requires three types of hydrolytic enzymes: (1) Endoglucanases which cut internal beta-1,4-glucosidic bonds; (2) Exocellobiohydrolases that cut the dissaccharide cellobiose from the non-reducing end of the cellulose polymer chain; (3) Beta-1,4-glucosidases which hydrolyze the cellobiose and other short cello-oligosaccharides to glucose.
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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/oj/1ojj_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.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=1ojj ConSurf].
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<div style="clear:both"></div>
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<div style="background-color:#fffaf0;">
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== Publication Abstract from PubMed ==
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The formation of glycoconjugates and oligosaccharides remains one of the most challenging chemical syntheses. Chemo-enzymatic routes using retaining glycosidases have been successfully harnessed but require tight kinetic or thermodynamic control. "Glycosynthases," specifically engineered glycosidases that catalyze the formation of glycosidic bonds from glycosyl donor and acceptor alcohol, are an emerging range of synthetic tools in which catalytic nucleophile mutants are harnessed together with glycosyl fluoride donors to generate powerful and versatile catalysts. Here we present the structural and kinetic dissection of the Humicola insolens Cel7B glycosynthases in which the nucleophile of the wild-type enzyme is mutated to alanine and serine (E197A and E197S). 3-D structures reveal the acceptor and donor subsites and the basis for substrate inhibition. Kinetic analysis shows that the E197S mutant is considerably more active than the corresponding alanine mutant due to a 40-fold increase in k(cat).
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==About this Structure==
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Anatomy of glycosynthesis: structure and kinetics of the Humicola insolens Cel7B E197A and E197S glycosynthase mutants.,Ducros VM, Tarling CA, Zechel DL, Brzozowski AM, Frandsen TP, von Ossowski I, Schulein M, Withers SG, Davies GJ Chem Biol. 2003 Jul;10(7):619-28. PMID:12890535<ref>PMID:12890535</ref>
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1OJJ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Humicola_insolens Humicola insolens] with NAG as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] Structure known Active Site: AC1. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1OJJ OCA].
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==Reference==
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From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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Anatomy of glycosynthesis: structure and kinetics of the Humicola insolens Cel7B E197A and E197S glycosynthase mutants., Ducros VM, Tarling CA, Zechel DL, Brzozowski AM, Frandsen TP, von Ossowski I, Schulein M, Withers SG, Davies GJ, Chem Biol. 2003 Jul;10(7):619-28. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=12890535 12890535]
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</div>
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[[Category: Cellulase]]
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<div class="pdbe-citations 1ojj" style="background-color:#fffaf0;"></div>
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[[Category: Humicola insolens]]
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[[Category: Single protein]]
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[[Category: Brzozowski, A.M.]]
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[[Category: Davies, G.J.]]
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[[Category: Ducros, V.M.A.]]
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[[Category: Frandsen, T.P.]]
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[[Category: Ossowski, I.Von.]]
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[[Category: Schulein, M.]]
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[[Category: Tarling, C.A.]]
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[[Category: Withers, S.G.]]
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[[Category: Zechel, D.L.]]
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[[Category: NAG]]
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[[Category: cellulose degradation]]
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[[Category: glycosynthase]]
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[[Category: hydrolase]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 16:56:45 2007''
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==See Also==
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*[[Glucanase 3D structures|Glucanase 3D structures]]
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== References ==
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<references/>
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__TOC__
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</StructureSection>
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[[Category: Humicola insolens]]
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[[Category: Large Structures]]
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[[Category: Brzozowski AM]]
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[[Category: Davies GJ]]
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[[Category: Ducros VM-A]]
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[[Category: Frandsen TP]]
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[[Category: Schulein M]]
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[[Category: Tarling CA]]
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[[Category: Von Ossowski I]]
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[[Category: Withers SG]]
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[[Category: Zechel DL]]

Current revision

Anatomy of glycosynthesis: Structure and kinetics of the Humicola insolens Cel7BE197A and E197S glycosynthase mutants

PDB ID 1ojj

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