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| | ==Ferredoxin-NADP(H) Reductase mutant with Ala 266 replaced by Tyr (A266Y) and residues 267-272 deleted.== | | ==Ferredoxin-NADP(H) Reductase mutant with Ala 266 replaced by Tyr (A266Y) and residues 267-272 deleted.== |
| - | <StructureSection load='4k1x' size='340' side='right' caption='[[4k1x]], [[Resolution|resolution]] 1.70Å' scene=''> | + | <StructureSection load='4k1x' size='340' side='right'caption='[[4k1x]], [[Resolution|resolution]] 1.70Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4k1x]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/"rhodonostoc_capsulatum"_molisch_1907 "rhodonostoc capsulatum" molisch 1907]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K1X OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4K1X FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4k1x]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Rhodobacter_capsulatus Rhodobacter capsulatus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4K1X OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4K1X FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.7Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2vni|2vni]], [[2bgj|2bgj]]</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">fpr ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1061 "Rhodonostoc capsulatum" Molisch 1907])</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=4k1x FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k1x OCA], [https://pdbe.org/4k1x PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4k1x RCSB], [https://www.ebi.ac.uk/pdbsum/4k1x PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4k1x ProSAT]</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=4k1x FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4k1x OCA], [http://pdbe.org/4k1x PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4k1x RCSB], [http://www.ebi.ac.uk/pdbsum/4k1x PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4k1x ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| - | <div style="background-color:#fffaf0;">
| + | == Function == |
| - | == Publication Abstract from PubMed == | + | [https://www.uniprot.org/uniprot/FENR_RHOCA FENR_RHOCA] Transports electrons between flavodoxin or ferredoxin and NADPH.<ref>PMID:14572660</ref> <ref>PMID:16128574</ref> <ref>PMID:24016470</ref> |
| - | To study the role of the mobile C-terminal extension present in bacterial class of plant type NADP(H):ferredoxin reductases during catalysis, we generated a series of mutants of the Rhodobacter capsulatus enzyme (RcFPR). Deletion of the six C-terminal amino acids beyond alanine 266 was combined with the replacement A266Y, emulating the structure present in plastidic versions of this flavoenzyme. Analysis of absorbance and fluorescence spectra suggests that deletion does not modify the general geometry of FAD itself, but increases exposure of the flavin to the solvent, prevents a productive geometry of FAD:NADP(H) complex and decreases the protein thermal stability. Although the replacement A266Y partially coats the isoalloxazine from solvent and slightly restores protein stability, this single change does not allow formation of active charge-transfer complexes commonly present in the wild-type FPR, probably due to restraints of C-terminus pliability. A proton exchange process is deduced from ITC measurements during coenzyme binding. All studied RcFPR variants display higher affinity for NADP+ than wild-type, evidencing the contribution of the C-terminus in tempering a non-productive strong (rigid) interaction with the coenzyme. The decreased catalytic rate parameters confirm that the hydride transfer from NADPH to the flavin ring is considerably hampered in the mutants. Although the involvement of the C-terminal extension from bacterial FPRs in stabilizing overall folding and bent-FAD geometry has been stated, the most relevant contributions to catalysis are modulation of coenzyme entrance and affinity, promotion of the optimal geometry of an active complex and supply of a proton acceptor acting during coenzyme binding.
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| - | The C-terminal extension of bacterial flavodoxin-reductases: Involvement in the hydride transfer mechanism from the coenzyme.,Bortolotti A, Sanchez-Azqueta A, Maya CM, Velazquez-Campoy A, Hermoso JA, Medina M, Cortez N Biochim Biophys Acta. 2013 Sep 6;1837(1):33-43. doi:, 10.1016/j.bbabio.2013.08.008. PMID:24016470<ref>PMID:24016470</ref>
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| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div> | + | |
| - | <div class="pdbe-citations 4k1x" style="background-color:#fffaf0;"></div> | + | |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Rhodonostoc capsulatum molisch 1907]] | + | [[Category: Large Structures]] |
| - | [[Category: Bortolotti, A]] | + | [[Category: Rhodobacter capsulatus]] |
| - | [[Category: Cortez, N]] | + | [[Category: Bortolotti A]] |
| - | [[Category: Hermoso, J A]] | + | [[Category: Cortez N]] |
| - | [[Category: Maya, C M]] | + | [[Category: Hermoso JA]] |
| - | [[Category: Sanchez-Azqueta, A]] | + | [[Category: Maya CM]] |
| - | [[Category: Velazquez-Campoy, A]] | + | [[Category: Sanchez-Azqueta A]] |
| - | [[Category: Nadp+ binding]]
| + | [[Category: Velazquez-Campoy A]] |
| - | [[Category: Oxidoreductase]]
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| - | [[Category: Reductase]]
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