4kqw: Difference between revisions
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==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=''> | ||
== 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> | |||
</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> | |||
<tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4kqx|4kqx]]</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> | |||
<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> | |||
<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://www.rcsb.org/pdb/explore.do?structureId=4kqw RCSB], [http://www.ebi.ac.uk/pdbsum/4kqw PDBsum]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== 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<ref>PMID:23776225</ref> | |||
== | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
</div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Ketol-acid reductoisomerase]] | [[Category: Ketol-acid reductoisomerase]] | ||
[[Category: Slackia exigua atcc 700122]] | [[Category: Slackia exigua atcc 700122]] | ||
[[Category: Arnold, F H | [[Category: Arnold, F H]] | ||
[[Category: Brinkmann-Chen, S | [[Category: Brinkmann-Chen, S]] | ||
[[Category: Brustad, E M | [[Category: Brustad, E M]] | ||
[[Category: Cahn, J K.B | [[Category: Cahn, J K.B]] | ||
[[Category: Flock, T | [[Category: Flock, T]] | ||
[[Category: Mcintosh, J A | [[Category: Mcintosh, J A]] | ||
[[Category: Meinhold, P | [[Category: Meinhold, P]] | ||
[[Category: Snow, C D | [[Category: Snow, C D]] | ||
[[Category: Zhang, L | [[Category: Zhang, L]] | ||
[[Category: Acetohydroxyacid isomeroreductase]] | [[Category: Acetohydroxyacid isomeroreductase]] | ||
[[Category: Oxidoreductase]] | [[Category: Oxidoreductase]] | ||
[[Category: Rossmann fold]] | [[Category: Rossmann fold]] | ||
