1. Spectroscopic Study of Substrate Binding to the Carbonmonoxy Form of Dehaloperoxidase from Amphitrite ornata.
- Author
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Karin Nienhaus, Pengchi Deng, Jennifer Belyea, Stefan Franzen, and G. Ulrich Nienhaus
- Subjects
- *
INFRARED spectroscopy , *SPECTRUM analysis , *CARBON monoxide , *DISINFECTION & disinfectants - Abstract
Dehaloperoxidase (DHP) is a globular heme enzyme found in the marine worm Amphitrite ornatathat can catalyze the dehalogenation of halophenols to the corresponding quinones by using hydrogen peroxide as a cosubstrate. Its three-dimensional fold is surprisingly similar to that of the oxygen storage protein myoglobin (Mb). A key structural feature common to both DHP and Mb is the existence of multiple conformations of the distal histidine. In DHP, the conformational flexibility may be involved in promotion of substrate and cosubstrate entry and exit. Here we have explored the dynamics of substrate binding in DHP using Fourier transform infrared spectroscopy and flash photolysis. A number of discrete conformations at the active site were identified from the appearance of multiple CO absorbance bands in the infrared region of the spectrum. Upon photolysis at cryogenic temperatures, the CO molecules are trapped at docking sites within the protein matrix, as inferred from the appearance of several photoproduct bands characteristic of each site. Substrate binding stabilizes the protein by ∼20 kJ/mol. The low yield of substrate-bound DHP at ambient temperature points toward a steric inhibition of substrate binding by carbon monoxide. [ABSTRACT FROM AUTHOR]
- Published
- 2006
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