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Solution Conformations and Dynamics of Substrate-Bound Cytochrome P450 MycG.
- Source :
-
Biochemistry [Biochemistry] 2017 May 30; Vol. 56 (21), pp. 2701-2714. Date of Electronic Publication: 2017 May 16. - Publication Year :
- 2017
-
Abstract
- MycG is a P450 monooxygenase that catalyzes the sequential hydroxylation and epoxidation of mycinamicin IV (M-IV), the last two steps in the biosynthesis of mycinamicin II, a macrolide antibiotic isolated from Micromonospora griseorubida. The crystal structure of MycG with M-IV bound was previously determined but showed the bound substrate in an orientation that did not rationalize the observed regiochemistry of M-IV hydroxylation. Nuclear magnetic resonance paramagnetic relaxation enhancements provided evidence of an orientation of M-IV in the MycG active site more compatible with the observed chemistry, but substrate-induced changes in the enzyme structure were not characterized. We now describe the use of amide <superscript>1</superscript> H- <superscript>15</superscript> N residual dipolar couplings as experimental restraints in solvated "soft annealing" molecular dynamics simulations to generate solution structural ensembles of M-IV-bound MycG. Chemical shift perturbations, hydrogen-deuterium exchange, and <superscript>15</superscript> N relaxation behavior provide insight into the dynamic and electronic perturbations in the MycG structure in response to M-IV binding. The solution and crystallographic structures are compared, and the possibility that the crystallographic orientation of bound M-IV represents an inhibitory mode is discussed.
Details
- Language :
- English
- ISSN :
- 1520-4995
- Volume :
- 56
- Issue :
- 21
- Database :
- MEDLINE
- Journal :
- Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 28488849
- Full Text :
- https://doi.org/10.1021/acs.biochem.7b00291