Analytical and Bioanalytical Chemistry

, Volume 399, Issue 6, pp 1999–2014 | Cite as

Ubiquinol formation in isolated photosynthetic reaction centres monitored by time-resolved differential FTIR in combination with 2D correlation spectroscopy and multivariate curve resolution

  • Alberto Mezzetti
  • Lionel Blanchet
  • Anna de Juan
  • Winfried Leibl
  • Cyril RuckebuschEmail author
Original Paper


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 photo-induced 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 (Q A /QA transition, first contribution), for a monoelectronic reduction of a secondary quinone QB (Q B /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.


Chemometrics Multivariate curve resolution 2D correlation spectroscopy Rapid-scan FTIR Purple bacteria 

Supplementary material

216_2010_4325_MOESM1_ESM.pdf (248 kb)
ESM 1 (PDF 247 KB)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Alberto Mezzetti
    • 1
    • 2
  • Lionel Blanchet
    • 3
  • Anna de Juan
    • 4
  • Winfried Leibl
    • 2
  • Cyril Ruckebusch
    • 1
    Email author
  1. 1.LASIR CNRS UMR 8516Université Lille1 Sciences et TechnologiesVilleneuve d’AscqFrance
  2. 2.LPB, Service de Bioénergétique, Biologie Structurale et Mécanismes, URA CNRS 2096, IBITec-S, CEA-SaclayGif-sur-Yvette cedexFrance
  3. 3.Institute for Molecules and Materials, Department of Analytical ChemistryRadboud University NijmegenNijmegenThe Netherlands
  4. 4.Department of Analytical ChemistryUniversitat de BarcelonaBarcelonaSpain

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