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| | ==Structure of a Lys-HCT mutant from Coffea canephora (Crystal form 1)== | | ==Structure of a Lys-HCT mutant from Coffea canephora (Crystal form 1)== |
| - | <StructureSection load='4g22' size='340' side='right' caption='[[4g22]], [[Resolution|resolution]] 1.70Å' scene=''> | + | <StructureSection load='4g22' size='340' side='right'caption='[[4g22]], [[Resolution|resolution]] 1.70Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4g22]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Cofca Cofca]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4G22 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4G22 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4g22]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Coffea_canephora Coffea canephora]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4G22 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4G22 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> | + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=CSO:S-HYDROXYCYSTEINE'>CSO</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> |
| - | <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CSO:S-HYDROXYCYSTEINE'>CSO</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=4g22 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4g22 OCA], [https://pdbe.org/4g22 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4g22 RCSB], [https://www.ebi.ac.uk/pdbsum/4g22 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4g22 ProSAT]</span></td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4g0b|4g0b]], [[4g2m|4g2m]]</td></tr>
| + | |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">HCT ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=49390 COFCA])</td></tr>
| + | |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Shikimate_O-hydroxycinnamoyltransferase Shikimate O-hydroxycinnamoyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.3.1.133 2.3.1.133] </span></td></tr>
| + | |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4g22 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4g22 OCA], [http://pdbe.org/4g22 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4g22 RCSB], [http://www.ebi.ac.uk/pdbsum/4g22 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4g22 ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/A4ZKE4_COFCA A4ZKE4_COFCA] |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Cofca]] | + | [[Category: Coffea canephora]] |
| - | [[Category: Shikimate O-hydroxycinnamoyltransferase]] | + | [[Category: Large Structures]] |
| - | [[Category: Lallemand, L A]] | + | [[Category: Lallemand LA]] |
| - | [[Category: McCarthy, A A]] | + | [[Category: McCarthy AA]] |
| - | [[Category: McCarthy, J G]] | + | [[Category: McCarthy JG]] |
| - | [[Category: Bahd superfamily]]
| + | |
| - | [[Category: Hydroxycinnamoyltransferase]]
| + | |
| - | [[Category: Transferase]]
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| Structural highlights
Function
A4ZKE4_COFCA
Publication Abstract from PubMed
Chlorogenic acids (CGA) are a group of phenolic secondary metabolites produced by certain plant species and an important component of coffee. The CGAs have been implicated in biotic and abiotic stress responses, while the related shikimate esters are key intermediates for lignin biosynthesis. Here, two hydroxycinnamoyl-CoA shikimate/ quinate hydroxycinnamoyl-transferases (HCT/HQT) from Coffea canephora were biochemically characterized. We show for the first time that, in vitro, HCT is capable of synthesizing the 3,5-O-dicaffeoylquinic acid diester (diCQA), a major constituent of the immature coffee grain. In order to further understand the substrate specificity and catalytic mechanism of the HCT/HQT, we performed structural and mutagnesis studies of HCT. The three-dimensional structure of a native HCT and a proteolytically stable lysine mutant enabled the identification of important residues involved in substrate specificity and catalysis. Site-directed mutagenesis confirmed the role of residues Leu-400 and Phe-402 in substrate specificity and His-153 and the Val-31 to Pro-37 loop in catalysis. In addition, the His-154-Asn mutant was observed to produce 4-fold more diCGAs compared to the native protein. These data provide the first structural characterization of a HCT and, in conjunction with the biochemical and mutagenesis studies presented here, delineate the underlying molecular level determinants for substrate specificity and catalysis. This work has potential applications in fine-tuning the levels of shikimate and quinate esters (CGAs including diCGAs), in different plant species in order to generate reduced or elevated levels of the desired target compounds.
A STRUCTURAL BASIS FOR THE BIOSYNTHESIS OF THE MAJOR CHLOROGENIC ACIDS FOUND IN COFFEE.,Lallemand LA, Zubieta C, Lee SG, Wang Y, Acajjaoui S, Timmins J, McSweeney S, Jez JM, McCarthy JG, McCarthy AA Plant Physiol. 2012 Jul 20. PMID:22822210[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Lallemand LA, Zubieta C, Lee SG, Wang Y, Acajjaoui S, Timmins J, McSweeney S, Jez JM, McCarthy JG, McCarthy AA. A STRUCTURAL BASIS FOR THE BIOSYNTHESIS OF THE MAJOR CHLOROGENIC ACIDS FOUND IN COFFEE. Plant Physiol. 2012 Jul 20. PMID:22822210 doi:10.1104/pp.112.202051
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