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| | ==The structure of the Slackia exigua KARI in complex with NADP== | | ==The structure of the Slackia exigua KARI in complex with NADP== |
| - | <StructureSection load='4kqw' size='340' side='right' caption='[[4kqw]], [[Resolution|resolution]] 1.39Å' scene=''> | + | <StructureSection load='4kqw' size='340' side='right'caption='[[4kqw]], [[Resolution|resolution]] 1.39Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4kqw]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Slackia_exigua_atcc_700122 Slackia exigua atcc 700122]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4KQW OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4KQW FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4kqw]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Slackia_exigua_ATCC_700122 Slackia exigua ATCC 700122]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4KQW OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4KQW FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</scene>, <scene name='pdbligand=TLA:L(+)-TARTARIC+ACID'>TLA</scene></td></tr> | + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NAP:NADP+NICOTINAMIDE-ADENINE-DINUCLEOTIDE+PHOSPHATE'>NAP</scene>, <scene name='pdbligand=TLA:L(+)-TARTARIC+ACID'>TLA</scene></td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4kqx|4kqx]]</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=4kqw FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4kqw OCA], [https://pdbe.org/4kqw PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4kqw RCSB], [https://www.ebi.ac.uk/pdbsum/4kqw PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4kqw ProSAT]</span></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">ilvC, HMPREF0762_00954 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=649764 Slackia exigua ATCC 700122])</td></tr>
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| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/Ketol-acid_reductoisomerase Ketol-acid reductoisomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.86 1.1.1.86] </span></td></tr>
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| - | <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=4kqw FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4kqw OCA], [http://pdbe.org/4kqw PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4kqw RCSB], [http://www.ebi.ac.uk/pdbsum/4kqw PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4kqw ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/ILVC_SLAES ILVC_SLAES] Involved in the biosynthesis of branched-chain amino acids (BCAA). Catalyzes an alkyl-migration followed by a ketol-acid reduction of (S)-2-acetolactate (S2AL) to yield (R)-2,3-dihydroxy-isovalerate. In the isomerase reaction, S2AL is rearranged via a Mg-dependent methyl migration to produce 3-hydroxy-3-methyl-2-ketobutyrate (HMKB). In the reductase reaction, this 2-ketoacid undergoes a metal-dependent reduction by NADPH to yield (R)-2,3-dihydroxy-isovalerate.[HAMAP-Rule:MF_00435]<ref>PMID:23776225</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | ==See Also== | | ==See Also== |
| - | *[[Ketol-acid reductoisomerase|Ketol-acid reductoisomerase]] | + | *[[Ketol-acid reductoisomerase 3D structures|Ketol-acid reductoisomerase 3D structures]] |
| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Ketol-acid reductoisomerase]] | + | [[Category: Large Structures]] |
| - | [[Category: Slackia exigua atcc 700122]] | + | [[Category: Slackia exigua ATCC 700122]] |
| - | [[Category: Arnold, F H]] | + | [[Category: Arnold FH]] |
| - | [[Category: Brinkmann-Chen, S]] | + | [[Category: Brinkmann-Chen S]] |
| - | [[Category: Brustad, E M]] | + | [[Category: Brustad EM]] |
| - | [[Category: Cahn, J K.B]] | + | [[Category: Cahn JKB]] |
| - | [[Category: Flock, T]] | + | [[Category: Flock T]] |
| - | [[Category: Mcintosh, J A]] | + | [[Category: Mcintosh JA]] |
| - | [[Category: Meinhold, P]] | + | [[Category: Meinhold P]] |
| - | [[Category: Snow, C D]] | + | [[Category: Snow CD]] |
| - | [[Category: Zhang, L]] | + | [[Category: Zhang L]] |
| - | [[Category: Acetohydroxyacid isomeroreductase]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| - | [[Category: Rossmann fold]]
| + | |
| Structural highlights
Function
ILVC_SLAES Involved in the biosynthesis of branched-chain amino acids (BCAA). Catalyzes an alkyl-migration followed by a ketol-acid reduction of (S)-2-acetolactate (S2AL) to yield (R)-2,3-dihydroxy-isovalerate. In the isomerase reaction, S2AL is rearranged via a Mg-dependent methyl migration to produce 3-hydroxy-3-methyl-2-ketobutyrate (HMKB). In the reductase reaction, this 2-ketoacid undergoes a metal-dependent reduction by NADPH to yield (R)-2,3-dihydroxy-isovalerate.[HAMAP-Rule:MF_00435][1]
Publication Abstract from PubMed
To date, efforts to switch the cofactor specificity of oxidoreductases from nicotinamide adenine dinucleotide phosphate (NADPH) to nicotinamide adenine dinucleotide (NADH) have been made on a case-by-case basis with varying degrees of success. Here we present a straightforward recipe for altering the cofactor specificity of a class of NADPH-dependent oxidoreductases, the ketol-acid reductoisomerases (KARIs). Combining previous results for an engineered NADH-dependent variant of Escherichia coli KARI with available KARI crystal structures and a comprehensive KARI-sequence alignment, we identified key cofactor specificity determinants and used this information to construct five KARIs with reversed cofactor preference. Additional directed evolution generated two enzymes having NADH-dependent catalytic efficiencies that are greater than the wild-type enzymes with NADPH. High-resolution structures of a wild-type/variant pair reveal the molecular basis of the cofactor switch.
General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH.,Brinkmann-Chen S, Flock T, Cahn JK, Snow CD, Brustad EM, McIntosh JA, Meinhold P, Zhang L, Arnold FH Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10946-51. doi:, 10.1073/pnas.1306073110. Epub 2013 Jun 17. PMID:23776225[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Brinkmann-Chen S, Flock T, Cahn JK, Snow CD, Brustad EM, McIntosh JA, Meinhold P, Zhang L, Arnold FH. General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH. Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10946-51. doi:, 10.1073/pnas.1306073110. Epub 2013 Jun 17. PMID:23776225 doi:10.1073/pnas.1306073110
- ↑ Brinkmann-Chen S, Flock T, Cahn JK, Snow CD, Brustad EM, McIntosh JA, Meinhold P, Zhang L, Arnold FH. General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH. Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10946-51. doi:, 10.1073/pnas.1306073110. Epub 2013 Jun 17. PMID:23776225 doi:10.1073/pnas.1306073110
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