Ubiquinol formation in isolated photosynthetic reaction centres monitored by time-resolved differential FTIR in combination with 2D correlation spectroscopy and multivariate curve resolution.
Two-dimensional correlation analysis was carried out in combination with multivariate curve resolution-alternating least squares (MCR-ALS) to analyse time-resolved infrared (IR) difference spectra probing photoinduced ubiquinol formation in detergent-isolated reaction centres from Rhodobacter sphaeroides. The dynamic 2D IR correlation spectra have not only allowed the determination of the concomitance or non-concomitance of different chemical events through known marker bands but also have helped identify new vibrational bands related to the complex series of photochemical and redox reactions. In particular, a strong positive band located at 1565 cm⁻¹ was found to be synchronous with the process of ubiquinol formation. In addition, a tailored MCR-ALS analysis was performed using a priori chemical knowledge of the system, in particular including the pure spectrum of one species obtained from an external measurement. Enhancing the MCR-ALS performance in this way in time-dependent processes is relevant, especially when other essential pieces of information, such as kinetic models, are unavailable. The results give evidence of four independent spectral contributions. Three of them show marker bands for a monoelectronic reduction of the primary quinone QA (QA⁻/QA transition, first contribution), for a monoelectronic reduction of a secondary quinone QB (QB⁻/QB transition, second contribution) and for ubiquinol formation (third contribution). The results obtained also confirm that a key rate-limiting factor is the slow ubiquinone and ubiquinol exchange among micelles, which strongly influences the kinetic profiles of the involved species.
Université Lille1 Sciences et Technologies, 59655 Villeneuve d'Ascq, France.
Alberto
A
Mezzetti
Lionel
L
Blanchet
Anna
A
de Juan
Winfried
W
Leibl
Cyril
C
Ruckebusch
D007700:Kinetics; D008956:Models, Chemical; D015999:Multivariate Analysis; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000737:chemistry* / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D013057:Spectrum Analysis; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
D045322:Photosynthetic Reaction Center Complex Proteins; D014451:Ubiquinone; C003741:ubiquinol