title: Low-frequency dynamics of Caldariomyces fumago chloroperoxidase probed by femtosecond coherence spectroscopy.
abstract: Ultrafast laser spectroscopy techniques are used to measure the low-frequency vibrational coherence spectra and nitric oxide rebinding kinetics of Caldariomyces fumago chloroperoxidase (CPO). Comparisons of the CPO coherence spectra with those of other heme species are made to gauge the protein-specific nature of the low-frequency spectra. The coherence spectrum of native CPO is dominated by a mode that appears near 32-33 cm(-1) at all excitation wavelengths, with a phase that is consistent with a ground-state Raman-excited vibrational wavepacket. On the basis of a normal coordinate structural decomposition (NSD) analysis, we assign this feature to the thiolate-bound heme doming mode. Spectral resolution of the probe pulse ("detuned" detection) reveals a mode at 349 cm(-1), which has been previously assigned using Raman spectroscopy to the Fe-S stretching mode of native CPO. The ferrous species displays a larger degree of spectral inhomogeneity than the ferric species, as reflected by multiple shoulders in the optical absorption spectra. The inhomogeneities are revealed by changes in the coherence spectra at different excitation wavelengths. The appearance of a mode close to 220 cm(-1) in the coherence spectrum of reduced CPO excited at 440 nm suggests that a subpopulation of five coordinated histidine-ligated hemes is present in the ferrous state at a physiologically relevant pH. A significant increase in the amplitude of the coherence signal is observed for the resonance with the 440 nm subpopulation. Kinetics measurements reveal that nitric oxide binding to ferric and ferrous CPO can be described as a single-exponential process, with rebinding time constants of 29.4 +/- 1 and 9.3 +/- 1 ps, respectively. This is very similar to results previously reported for nitric oxide binding to horseradish peroxidase.
authors.affiliation: Department of Physics and Center for Interdisciplinary Research on Complex Systems, Northeastern University, Boston, Massachusetts 02115, USA.
authors.forename: Flaviu
authors.initials: F
authors.lastname: Gruia
authors.forename: Dan
authors.initials: D
authors.lastname: Ionascu
authors.forename: Minoru
authors.initials: M
authors.lastname: Kubo
authors.forename: Xiong
authors.initials: X
authors.lastname: Ye
authors.forename: John
authors.initials: J
authors.lastname: Dawson
authors.forename: Robert L
authors.initials: RL
authors.lastname: Osborne
authors.forename: S G
authors.initials: SG
authors.lastname: Sligar
authors.forename: Ilia
authors.initials: I
authors.lastname: Denisov
authors.forename: Aditi
authors.initials: A
authors.lastname: Das
authors.forename: T L
authors.initials: TL
authors.lastname: Poulos
authors.forename: James
authors.initials: J
authors.lastname: Terner
authors.forename: Paul M
authors.initials: PM
authors.lastname: Champion
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D001665:Binding Sites; D002711:Chloride Peroxidase / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D013057:Spectrum Analysis; D013059:Spectrum Analysis, Raman; D013997:Time Factors
chemical_list: D009569:Nitric Oxide; D002711:Chloride Peroxidase