Photosynthesis Research

, Volume 142, Issue 3, pp 307–319 | Cite as

Near-infrared in vitro measurements of photosystem I cofactors and electron-transfer partners with a recently developed spectrophotometer

  • Pierre SétifEmail author
  • Alain Boussac
  • Anja Krieger-Liszkay
Original article


A kinetic-LED-array-spectrophotometer (Klas) was recently developed for measuring in vivo redox changes of P700, plastocyanin (PCy), and ferredoxin (Fd) in the near-infrared (NIR). This spectrophotometer is used in the present work for in vitro light-induced measurements with various combinations of photosystem I (PSI) from tobacco and two different cyanobacteria, spinach plastocyanin, cyanobacterial cytochrome c6 (cyt. c6), and Fd. It is shown that cyt. c6 oxidation contributes to the NIR absorption changes. The reduction of (FAFB), the terminal electron acceptor of PSI, was also observed and the shape of the (FAFB) NIR difference spectrum is similar to that of Fd. The NIR difference spectra of the electron-transfer cofactors were compared between different organisms and to those previously measured in vivo, whereas the relative absorption coefficients of all cofactors were determined by using single PSI turnover conditions. Thus, the (840 nm minus 965 nm) extinction coefficients of the light-induced species (oxidized minus reduced for PC and cyt. c6, reduced minus oxidized for (FAFB), and Fd) were determined with values of 0.207 ± 0.004, – 0.033 ± 0.006, – 0.036 ± 0.008, and – 0.021 ± 0.005 for PCy, cyt. c6, (FAFB) (single reduction), and Fd, respectively, by taking a reference value of + 1 for P700+. The fact that the NIR P700 coefficient is larger than that of PCy and much larger than that of other contributing species, combined with the observed variability in the NIR P700 spectral shape, emphasizes that deconvolution of NIR signals into different components requires a very precise determination of the P700 spectrum.


P700 Plastocyanin Cytochrome c6 Photosystem I terminal acceptor Ferredoxin Infrared spectral deconvolution 



A. K.-L. and P.S. most gratefully thank Dr. C. Klughammer for his help in installing the Klas-NIR spectrophotometer and in using the associated software. They also thank the LabEx Saclay Plant Sciences-SPS (ANR-10-LABX-0040-SPS) for its financial support for the acquisition of the Klas-NIR spectrophotometer. This work was also partially supported by the French Infrastructure for Integrated Structural Biology (FRISBI, ANR-10-INBS-05a) and by a grant from the Agence Nationale de la Recherche (RECYFUEL project, ANR-16-CE05-0026).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11120_2019_665_MOESM1_ESM.docx (513 kb)
Supplementary material 1 (DOCX 513 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, Université Paris-SaclayGif-Sur-Yvette CedexFrance

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