title: Spectroscopic observation of the intramolecular electron transfer in the photoactivation processes of nitrile hydratase.
abstract: The photoactivation phenomena of the photosensitive enzyme nitrile hydratase (NHase) was studied by various spectroscopic methods. We have already shown that the photoactivation of NHase accompanies oxidation of an iron atom in the NHase [Honda et al. (1992) FEBS Lett. 301, 177-180]. From the results obtained in the present study by absorption, action, and fluorescence spectra, we show that the chromophore responsible for the photoactivation process is the iron complex, and the tryptophan residues in NHase induce the oxidation of the iron atom via an energy-transfer process. The nanosecond flash photolysis experiment revealed that this photoactivation process is completed within 50 ns, which suggests that the changes observable in the absorption spectra originate from an intramolecular electron transfer occurring from an electronically excited state. Also the role of a stabilizing reagent, namely, n-butyric acid (BA), was investigated using the above methods, which revealed that BA, besides its stabilizing effect, contributes to the increase in apparent photoactivation rate.
authors.affiliation: Frontier Research Program, Institute of Physical and Chemical Research (RIKEN), Saitama.
authors.forename: J
authors.initials: J
authors.lastname: Honda
authors.forename: H
authors.initials: H
authors.lastname: Kandori
authors.forename: T
authors.initials: T
authors.lastname: Okada
authors.forename: T
authors.initials: T
authors.lastname: Nagamune
authors.forename: Y
authors.initials: Y
authors.lastname: Shichida
authors.forename: H
authors.initials: H
authors.lastname: Sasabe
authors.forename: I
authors.initials: I
authors.lastname: Endo
mesh_terms: D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D004579:Electron Transport; D004789:Enzyme Activation / Q000528:radiation effects; D006836:Hydro-Lyases / Q000737:chemistry / Q000378:metabolism*; D008027:Light; D010777:Photochemistry; D012240:Rhodococcus / Q000201:enzymology; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
chemical_list: D002087:Butyrates; D020148:Butyric Acid; D006836:Hydro-Lyases; C048831:nitrile hydratase