title: Coupled flexibility change in cytochrome P450cam substrate binding determined by neutron scattering, NMR, and molecular dynamics simulation.
abstract: Neutron scattering and nuclear magnetic resonance relaxation experiments are combined with molecular dynamics (MD) simulations in a novel, to our knowledge, approach to investigate the change in internal dynamics on substrate (camphor) binding to a protein (cytochrome P450cam). The MD simulations agree well with both the neutron scattering, which furnishes information on global flexibility, and the nuclear magnetic resonance data, which provides residue-specific order parameters. Decreased fluctuations are seen in the camphor-bound form using all three techniques, dominated by changes in specific regions of the protein. The combined experimental and simulation results permit a detailed description of the dynamical change, which involves modifications in the coupling between the dominant regions and concomitant substrate access channel closing, via specific salt-bridge, hydrogen-bonding, and hydrophobic interactions. The work demonstrates how the combination of complementary experimental spectroscopies with MD simulation can provide an in-depth description of functional dynamical protein changes.
authors.affiliation: University of Tennessee/Oak Ridge National Laboratory Center for Molecular Biophysics, Oak Ridge National Laboratory, Oak Ridge, TN, USA.
authors.forename: Yinglong
authors.initials: Y
authors.lastname: Miao
authors.forename: Zheng
authors.initials: Z
authors.lastname: Yi
authors.forename: Carey
authors.initials: C
authors.lastname: Cantrell
authors.forename: Dennis C
authors.initials: DC
authors.lastname: Glass
authors.forename: Jerome
authors.initials: J
authors.lastname: Baudry
authors.forename: Nitin
authors.initials: N
authors.lastname: Jain
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Smith
mesh_terms: D019475:Camphor 5-Monooxygenase / Q000737:chemistry*; D018360:Crystallography, X-Ray; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D009682:Magnetic Resonance Spectroscopy; D056004:Molecular Dynamics Simulation; D033363:Neutron Diffraction; D016958:Pseudomonas putida / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D019475:Camphor 5-Monooxygenase