4J6D

The 2.4 A crystal structure of CYP154C5 from Nocardia farcinica in complex with testosterone

Summary for 4J6D

• Entry DOI: 10.2210/pdb4j6d/pdb
• Related: 4J6B 4J6C 4JBT
• Descriptor: Cytochrome P450 monooxygenase, PROTOPORPHYRIN IX CONTAINING FE, MAGNESIUM ION, ... (7 entities in total)
• Functional Keywords: cytochrom p450, steroid hydroxylating monooxygenase, steroid binding, oxidoreductase-substrate complex, oxidoreductase/substrate
• Biological source: Nocardia farcinica
• Total number of polymer chains: 2
• Total formula weight: 92874.35

Authors

Herzog, K.,Hoffmann, K.M. (deposition date: 2013-02-11, release date: 2014-03-05, Last modification date: 2023-09-20)

Primary citation

Herzog, K.,Bracco, P.,Onoda, A.,Hayashi, T.,Hoffmann, K.,Schallmey, A.
Enzyme-substrate complex structures of CYP154C5 shed light on its mode of highly selective steroid hydroxylation.
Acta Crystallogr.,Sect.D, 70:2875-2889, 2014

PubMed Abstract: 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 16α-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 Å. 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.

PubMed: 25372679
DOI: 10.1107/S1399004714019129

Experimental method

X-RAY DIFFRACTION (2.4 Å)