4ax6: Difference between revisions
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<StructureSection load='4ax6' size='340' side='right'caption='[[4ax6]], [[Resolution|resolution]] 2.30Å' scene=''> | <StructureSection load='4ax6' size='340' side='right'caption='[[4ax6]], [[Resolution|resolution]] 2.30Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[4ax6]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4AX6 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4AX6 FirstGlance]. <br> | <table><tr><td colspan='2'>[[4ax6]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Trichoderma_reesei Trichoderma reesei]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4AX6 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4AX6 FirstGlance]. <br> | ||
</td></tr><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=MAN:ALPHA-D-MANNOSE'>MAN</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=UWU:6-CHLORANYL-7-OXIDANYL-4-PHENYL-CHROMEN-2-ONE'>UWU</scene | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.3Å</td></tr> | ||
<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=MAN:ALPHA-D-MANNOSE'>MAN</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=PRD_900005:beta-cellobiose'>PRD_900005</scene>, <scene name='pdbligand=UWU:6-CHLORANYL-7-OXIDANYL-4-PHENYL-CHROMEN-2-ONE'>UWU</scene></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=4ax6 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4ax6 OCA], [https://pdbe.org/4ax6 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4ax6 RCSB], [https://www.ebi.ac.uk/pdbsum/4ax6 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4ax6 ProSAT]</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=4ax6 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4ax6 OCA], [https://pdbe.org/4ax6 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4ax6 RCSB], [https://www.ebi.ac.uk/pdbsum/4ax6 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4ax6 ProSAT]</span></td></tr> | ||
</table> | </table> | ||
== Function == | == Function == | ||
[https://www.uniprot.org/uniprot/GUX2_HYPJE GUX2_HYPJE] 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. | |||
<div style="background-color:#fffaf0;"> | <div style="background-color:#fffaf0;"> | ||
== Publication Abstract from PubMed == | == Publication Abstract from PubMed == | ||
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</StructureSection> | </StructureSection> | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Hansson | [[Category: Trichoderma reesei]] | ||
[[Category: Ishida | [[Category: Hansson H]] | ||
[[Category: Nerinckx | [[Category: Ishida T]] | ||
[[Category: Piens | [[Category: Nerinckx W]] | ||
[[Category: Sandgren | [[Category: Piens K]] | ||
[[Category: Stahlberg | [[Category: Sandgren M]] | ||
[[Category: Wu | [[Category: Stahlberg J]] | ||
[[Category: Wu M]] | |||
Latest revision as of 14:37, 20 December 2023
HYPOCREA JECORINA CEL6A D221A MUTANT SOAKED WITH 6-CHLORO-4- PHENYLUMBELLIFERYL-BETA-CELLOBIOSIDEHYPOCREA JECORINA CEL6A D221A MUTANT SOAKED WITH 6-CHLORO-4- PHENYLUMBELLIFERYL-BETA-CELLOBIOSIDE
Structural highlights
FunctionGUX2_HYPJE 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. Publication Abstract from PubMedMethylumbelliferyl-beta-cellobioside (MUF-G2) is a convenient fluorogenic substrate for certain beta-glycoside hydrolases (GH). However, hydrolysis of the aglycone is poor with GH family 6 enzymes (GH6), despite strong binding. Prediction of the orientation of the aglycone of MUF-G2 in the +1 subsite of Hypocrea jecorina Cel6A by automated docking suggested umbelliferyl modifications at C4 and C6 for improved recognition. Four modified umbelliferyl-beta-cellobiosides [6-chloro-4-methyl- (ClMUF); 6-chloro-4-trifluoromethyl- (ClF3MUF); 4-phenyl- (PhUF); 6-chloro-4-phenyl- (ClPhUF)] were synthesized and tested with GH6, GH7, GH9, GH5 and GH45 cellulases. Indeed the rate of aglycone release by H. jecorina Cel6A was 10-150 times higher than with MUF-G2, although it was still three orders of magnitude lower than with H. jecorina Cel7B. The 4-phenyl substitution drastically reduced the fluorescence intensity of the free aglycone, while ClMUF-G2 could be used for determination of k(cat) and K(M) for H. jecorina Cel6A and Thermobifida fusca Cel6A. Crystal structures of H. jecorina Cel6A D221A mutant soaked with the MUF-, ClMUF- and ClPhUF-beta-cellobioside substrates show that the modifications turned the umbelliferyl group 'upside down', with the glycosidic bond better positioned for protonation than with MUF-G2. DATABASE: Structural data have been submitted to the Protein Data Bank under accession numbers pdb 4AU0, 4AX7, 4AX6 STRUCTURED DIGITAL ABSTRACT: * http://mint.bio.uniroma2.it/mint/search/interaction.do?interactionAc=MINT-7260296 * Cel6A and Cel6A bind by x-ray crystallography (View Interaction: 1, 2). Rational design, synthesis, evaluation and enzyme-substrate structures of improved fluorogenic substrates for family 6 glycoside hydrolases.,Wu M, Nerinckx W, Piens K, Ishida T, Hansson H, Sandgren M, Stahlberg J FEBS J. 2013 Jan;280(1):184-98. doi: 10.1111/febs.12060. Epub 2012 Dec 7. PMID:23137336[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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