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Dynamics Underlying Cytochrome P450cam Regioselectivity via 2D IR Spectroscopy
Author(s) -
Thielges Megan,
Basom Edward
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.762.9
Subject(s) - regioselectivity , chemistry , molecular dynamics , dynamics (music) , cytochrome , protein dynamics , substrate (aquarium) , active site , spectroscopy , stereochemistry , cytochrome p450 , computational chemistry , enzyme , chemical physics , biophysics , biochemistry , biology , physics , catalysis , quantum mechanics , acoustics , ecology
Conformational heterogeneity and dynamics likely contribute to the remarkable activity of enzymes, but are challenging to characterize experimentally. We applied linear and 2D IR spectroscopy to compare the conformational heterogeneity and dynamics of wild‐type and Y96F cytochrome P450cam‐CO bound to substrates that are hydroxylated with varying regioselectively. Our results suggest that the binding of different substrates to P450cam variably stabilizes the active site into two distinct states associated with different levels of regioselectivity. P450cam regioselectivity appears to be unrelated to the average electrostatic environment in the protein‐substrate complexes; instead, regioselectivity is correlated with conformational heterogeneity and dynamics within the populated conformations, supporting that fast dynamics contribute to the activity of this enzyme. We are extending the studies to measure conformations and dynamics throughout this and other proteins via the selective incorporations of IR‐active probe groups with spectrally resolved frequencies as an approach to the measurement of dynamics of proteins with high temporal and spatial resolution.