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.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Miao Y,Yi Z,Cantrell C,Glass DC,Baudry J,Jain N,Smith JC

doi

10.1016/j.bpj.2012.10.013

subject

Has Abstract

pub_date

2012-11-21 00:00:00

pages

2167-76

issue

10

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(12)01117-4

journal_volume

103

pub_type

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