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Time-resolved resonance Raman elucidation of the pathway for dioxygen reduction by cytochrome c oxidase

Journal Article · · Journal of the American Chemical Society; (United States)
DOI:https://doi.org/10.1021/ja00072a002· OSTI ID:5115096
; ; ;  [1]; ;  [2];  [3]
  1. Graduate Univ. of Advanced Studies, Okazaki (Japan)
  2. Himeji Inst. of Technology, Hyogo (Japan)
  3. Argonne National Lab., IL (United States)

Time-resolved resonance Raman (RR) spectra were observed for catalytic intermediates of bovine cytochrome c oxidase at room temperature by using an Artificial Cardiovascular System for Enzymatic Reactions and Raman/Absorption Simultaneous Measurement Device. The isotope-frequency-shift data using an asymmetrically labeled dioxygen, [sup 16]O[sup 18]O, established that the primary intermediate (compound A) is an end-on-type dioxygen adduct of cytochrome a[sub 3]. Higher resolution RR experiments revealed that the approximately 800-cm[sup [minus]1] band assigned previously to the Fe[sup TV]= O stretching mode of the subsequent intermediate (compound B) was actually composed of two bands that behaved differently upon deuteration. The 804/764-cm[sup [minus]1] pair for the [sup 16]O[sub 2]/[sup 18]O[sub 2] derivatives were insensitive to deuteration, and these bands became noticeably stronger at higher temperatures. The present experiments have conclusively established that the main pathway for dioxygen reduction by cytochrome c oxidase is Fe[sup II]-O[sub 2] [yields] Fe[sup III]-O-O-H [yields] Fe[sup IV]=O [yields] Fe[sup III]-OH and that their key bands for the [sup 16]O[sub 2]/[sup 18]O[sub 2] pair are at 571/544, 785/751, 804/764, and 450/425 cm[sup [minus]1], respectively. 79 refs., 8 figs., 3 tabs.

DOE Contract Number:
W-31109-ENG-38
OSTI ID:
5115096
Journal Information:
Journal of the American Chemical Society; (United States), Journal Name: Journal of the American Chemical Society; (United States) Vol. 115:19; ISSN JACSAT; ISSN 0002-7863
Country of Publication:
United States
Language:
English