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Article Dans Une Revue Journal of Raman Spectroscopy Année : 2011

Sub-picosecond Raman spectrometer for time-resolved studies of structural dynamics in heme proteins

Résumé

We describe a pump-probe Raman spectrometer based on a femtosecond Ti:sapphire laser, an optical parametric generator and two optical parametric amplifiers for time-resolved studies, with emphasis on the structural dynamics in heme proteins. The system provides a 100-fs pump pulse tunable in the range 500-600 nm and a transform-limited sub-picosecond probe pulse tunable in the range 390-450 nm. The spectrometer has spectral (25 cm(-1)) and temporal (similar to 0.7 ps) resolutions which constitute an effective compromise for identifying transient heme protein species and for following their structural evolution by spontaneous Raman scattering in the time range 0.5 ps to 2 ns. This apparatus was applied to time-resolved studies of a broad range of heme proteins, monitoring the primary dynamics of photoinduced heme coordination state and structural changes, its interaction with protein side-chains and diatomic gaseous ligands, as well as heme vibrational cooling. The treatment of transient Raman spectra is described in detail, and the advantages and shortcomings of spontaneous resonance Raman spectroscopy for ultrafast heme proteins studies are discussed. We demonstrate the efficiency of the constructed spectrometer by measuring Raman spectra in the sub-picosecond and picosecond time ranges for the oxygen-storage heme protein myoglobin and for the oxygen-sensor heme protein FixLH in interaction with the diatomic gaseous ligands CO, NO, and O-2. Copyright (C) 2010 John Wiley and Sons, Ltd.

Dates et versions

hal-00804600 , version 1 (25-03-2013)

Identifiants

Citer

Sergei G. Kruglik, Jean-Christophe Lambry, Jean-Louis Martin, Marten H. Vos, Michel Négrerie. Sub-picosecond Raman spectrometer for time-resolved studies of structural dynamics in heme proteins. Journal of Raman Spectroscopy, 2011, 42 (3), pp.265. ⟨10.1002/jrs.2685⟩. ⟨hal-00804600⟩
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