4j6b: Difference between revisions
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<StructureSection load='4j6b' size='340' side='right' caption='[[4j6b]], [[Resolution|resolution]] 2.20Å' scene=''> | <StructureSection load='4j6b' size='340' side='right' caption='[[4j6b]], [[Resolution|resolution]] 2.20Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[4j6b]] is a 2 chain structure | <table><tr><td colspan='2'>[[4j6b]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4J6B OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4J6B FirstGlance]. <br> | ||
</td></tr><tr><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=FMT:FORMIC+ACID'>FMT</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=PLO:(3BETA)-3-HYDROXYPREGN-5-EN-20-ONE'>PLO</scene>< | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=FMT:FORMIC+ACID'>FMT</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=PLO:(3BETA)-3-HYDROXYPREGN-5-EN-20-ONE'>PLO</scene></td></tr> | ||
<tr><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4j6c|4j6c]] | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4j6c|4j6c]], [[4jbt|4jbt]]</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=4j6b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4j6b OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4j6b RCSB], [http://www.ebi.ac.uk/pdbsum/4j6b PDBsum]</span></td></tr> | |||
<tr><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4j6b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4j6b OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4j6b RCSB], [http://www.ebi.ac.uk/pdbsum/4j6b PDBsum]</span></td></tr> | </table> | ||
<table> | <div style="background-color:#fffaf0;"> | ||
== Publication Abstract from PubMed == | |||
CYP154C5 from Nocardia farcinica is a bacterial cytochrome P450 monooxygenase active on steroid molecules. The enzyme has recently been shown to exhibit exclusive regioselectivity and stereoselectivity in the conversion of various pregnans and androstans, yielding 16alpha-hydroxylated steroid products. This makes the enzyme an attractive candidate for industrial application in steroid hormone synthesis. Here, crystal structures of CYP154C5 in complex with four different steroid molecules were solved at resolutions of up to 1.9 A. These are the first reported P450 structures from the CYP154 family in complex with a substrate. The active site of CYP154C5 forms a flattened hydrophobic channel with two opposing polar regions, perfectly resembling the size and polarity distribution of the steroids and thus resulting in highly specific steroid binding with Kd values in the range 10-100 nM. Key enzyme-substrate interactions were identified that accounted for the exclusive regioselectivity and stereoselectivity of the enzyme. Additionally, comparison of the four CYP154C5-steroid structures revealed distinct structural differences, explaining the observed variations in kinetic data obtained for this P450 with the steroids pregnenolone, dehydroepiandrosterone, progesterone, androstenedione, testosterone and nandrolone. This will facilitate the generation of variants with improved activity or altered selectivity in the future by means of protein engineering. | |||
Enzyme-substrate complex structures of CYP154C5 shed light on its mode of highly selective steroid hydroxylation.,Herzog K, Bracco P, Onoda A, Hayashi T, Hoffmann K, Schallmey A Acta Crystallogr D Biol Crystallogr. 2014 Nov;70(Pt 11):2875-89. doi:, 10.1107/S1399004714019129. Epub 2014 Oct 16. PMID:25372679<ref>PMID:25372679</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Herzog, K]] | |||
[[Category: Herzog, K | [[Category: Hoffmann, K M]] | ||
[[Category: Hoffmann, K M | |||
[[Category: Cytochrom p450]] | [[Category: Cytochrom p450]] | ||
[[Category: Oxidoreductase-substrate complex]] | [[Category: Oxidoreductase-substrate complex]] | ||
[[Category: Steroid binding]] | [[Category: Steroid binding]] | ||
[[Category: Steroid hydroxylating monooxygenase]] | [[Category: Steroid hydroxylating monooxygenase]] | ||
Revision as of 10:36, 12 November 2014
The 2.2 A crystal structure of CYP154C5 from Nocardia farcinica in complex with pregnenoloneThe 2.2 A crystal structure of CYP154C5 from Nocardia farcinica in complex with pregnenolone
Structural highlights
Publication Abstract from PubMedCYP154C5 from Nocardia farcinica is a bacterial cytochrome P450 monooxygenase active on steroid molecules. The enzyme has recently been shown to exhibit exclusive regioselectivity and stereoselectivity in the conversion of various pregnans and androstans, yielding 16alpha-hydroxylated steroid products. This makes the enzyme an attractive candidate for industrial application in steroid hormone synthesis. Here, crystal structures of CYP154C5 in complex with four different steroid molecules were solved at resolutions of up to 1.9 A. These are the first reported P450 structures from the CYP154 family in complex with a substrate. The active site of CYP154C5 forms a flattened hydrophobic channel with two opposing polar regions, perfectly resembling the size and polarity distribution of the steroids and thus resulting in highly specific steroid binding with Kd values in the range 10-100 nM. Key enzyme-substrate interactions were identified that accounted for the exclusive regioselectivity and stereoselectivity of the enzyme. Additionally, comparison of the four CYP154C5-steroid structures revealed distinct structural differences, explaining the observed variations in kinetic data obtained for this P450 with the steroids pregnenolone, dehydroepiandrosterone, progesterone, androstenedione, testosterone and nandrolone. This will facilitate the generation of variants with improved activity or altered selectivity in the future by means of protein engineering. Enzyme-substrate complex structures of CYP154C5 shed light on its mode of highly selective steroid hydroxylation.,Herzog K, Bracco P, Onoda A, Hayashi T, Hoffmann K, Schallmey A Acta Crystallogr D Biol Crystallogr. 2014 Nov;70(Pt 11):2875-89. doi:, 10.1107/S1399004714019129. Epub 2014 Oct 16. PMID:25372679[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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