title: Effects of crystallization on the heme-carbon monoxide moiety of bovine heart cytochrome c oxidase carbonyl.
abstract: Cytochrome c oxidase isolated from bovine heart was crystallized in the fully reduced carbon monoxide (CO)-bound form. To evaluate the structure of the O2 reaction site in crystals and in solution, the bound C-O stretch infrared band in protein crystals was compared with the band for protein solution. In solution, the C-O stretch band could be deconvoluted into two extremely narrow bands, one at 1963.6 cm-1 with delta v1/2 = 3.4 cm-1 of 60% Gaussian/40% Lorentzian character represented 86% of the total band area and the other at 1960.3 cm-1 with delta v1/2 = 3.0 cm-1 of 47% Gaussian/53% Lorentzian character represented 14% of the total band area. The crystals exhibited two deconvoluted C-O infrared bands having very similar band parameters with those in solution. These findings support the presence of two structurally similar conformers in both crystals and solution. Thus crystallization of this enzyme does not affect the structure at the CO-binding site to as great extent as has been noted for myoglobin and hemoglobin carbonyls, indicating that the active (CO- or O2-binding) site of cytochrome c oxidase must be conformationally very stable and highly ordered compared to other hemoproteins such as hemoglobin.
authors.affiliation: Department of Life Science, Faculty of Science, Himeji Institute of Technology, Hyogo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Tsubaki
authors.forename: K
authors.initials: K
authors.lastname: Shinzawa
authors.forename: S
authors.initials: S
authors.lastname: Yoshikawa
mesh_terms: D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002248:Carbon Monoxide / Q000737:chemistry; D002417:Cattle; D003460:Crystallization; D003576:Electron Transport Complex IV / Q000737:chemistry*; D006418:Heme / Q000737:chemistry; D015394:Molecular Structure; D009206:Myocardium / Q000201:enzymology; D012996:Solutions; D013055:Spectrophotometry, Infrared
chemical_list: D012996:Solutions; D006418:Heme; D002248:Carbon Monoxide; D003576:Electron Transport Complex IV