6LY4
The crystal structure of the BM3 mutant LG-23 in complex with testosterone
Summary for 6LY4
| Entry DOI | 10.2210/pdb6ly4/pdb |
| Descriptor | Bifunctional cytochrome P450/NADPH--P450 reductase, TESTOSTERONE, IMIDAZOLE, ... (6 entities in total) |
| Functional Keywords | p450-bm3 mutant, regio- and stereoselectivity, hydroxylation, testosterone, oxidoreductase |
| Biological source | Bacillus megaterium (strain ATCC 14581 / DSM 32 / JCM 2506 / NBRC 15308 / NCIMB 9376 / NCTC 10342 / VKM B-512) |
| Total number of polymer chains | 1 |
| Total formula weight | 56800.28 |
| Authors | Peng, Y.,Chen, J.,Zhou, J.,Li, A.,ReetZ, M.T. (deposition date: 2020-02-13, release date: 2020-04-29, Last modification date: 2023-11-29) |
| Primary citation | Li, A.,Acevedo-Rocha, C.G.,D'Amore, L.,Chen, J.,Peng, Y.,Garcia-Borras, M.,Gao, C.,Zhu, J.,Rickerby, H.,Osuna, S.,Zhou, J.,Reetz, M.T. Regio- and Stereoselective Steroid Hydroxylation at C7 by Cytochrome P450 Monooxygenase Mutants. Angew.Chem.Int.Ed.Engl., 59:12499-12505, 2020 Cited by PubMed Abstract: Steroidal C7β alcohols and their respective esters have shown significant promise as neuroprotective and anti-inflammatory agents to treat chronic neuronal damage like stroke, brain trauma, and cerebral ischemia. Since C7 is spatially far away from any functional groups that could direct C-H activation, these transformations are not readily accessible using modern synthetic organic techniques. Reported here are P450-BM3 mutants that catalyze the oxidative hydroxylation of six different steroids with pronounced C7 regioselectivities and β stereoselectivities, as well as high activities. These challenging transformations were achieved by a focused mutagenesis strategy and application of a novel technology for protein library construction based on DNA assembly and USER (Uracil-Specific Excision Reagent) cloning. Upscaling reactions enabled the purification of the respective steroidal alcohols in moderate to excellent yields. The high-resolution X-ray structure and molecular dynamics simulations of the best mutant unveil the origin of regio- and stereoselectivity. PubMed: 32243054DOI: 10.1002/anie.202003139 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.68 Å) |
Structure validation
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