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Rapid-Scan Fourier Transform Infrared Difference Spectroscopy with Two-Dimensional Correlation Analysis to Show the Build-Up of Light-Adapted States in Bacterial Photosynthetic Reaction Centers

Authors: Mezzetti A.; Malferrari M.; Venturoli G.; Francia F.; Leibl W.; Noda I.;

Rapid-Scan Fourier Transform Infrared Difference Spectroscopy with Two-Dimensional Correlation Analysis to Show the Build-Up of Light-Adapted States in Bacterial Photosynthetic Reaction Centers

Abstract

Time-resolved, rapid-scan Fourier transform infrared (FT-IR) difference spectra have been recorded upon illumination on photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides under fixed hydration conditions (relative humidity = 76%). Two different illumination schemes were adopted. Whereas the use of a laser flash (duration: 7 ns) made it possible to follow the kinetics of recombination of the light-induced state P + Q A – to the neutral state PQ A , the use of a 20.5 s continuous light from a lamp made it possible to follow both the build-up of a steady-state P + Q A – population and its decay to PQ A . Comparison between P + Q A – /PQ A FT-IR difference spectra obtained under (or 650 ms after) continuous illumination and obtained after one laser flash show small but meaningful differences, reflecting structural changes in the light-adapted state produced by the 20.5 s period of illumination. These differences are strikingly similar to those observed when comparing FT-IR difference spectra reflecting charge separation in photosystem II in light-adapted states and non-light-adapted states (c.f. Sipka et al., “Light-Adapted Charge-Separated State of Photosystem II: Structural and Functional Dynamics of the Closed Reaction Center”. Plant Cell. 2021. 33(4): 1286–1302). Two-dimensional correlation spectroscopy analysis revealed that in all the observed series of time-resolved FT-IR difference spectra (under illumination, after illumination, and after a laser flash), marker bands at 1749, 1716, and 1668 cm –1 all evolve synchronously, demonstrating that electron transfer reactions and protein backbone response (at least the one reflected by the 1668 cm –1 band) are strongly correlated. Conversely, for spectra under and after continuous illumination, many asynchronicities are observed for (still unassigned) bands throughout the whole 1740–1200 cm –1 region, reflecting a more complicated molecular scenario in the RC upon build-up of the light-adapted state and during its relaxation to the resting neutral state.

Keywords

Light, Spectroscopy, Fourier Transform Infrared, Photosynthetic Reaction Center Complex Proteins, Rhodobacter sphaeroides, 2D-COS; FT-IR; FT-IR difference spectroscopy; Time-resolved infrared spectroscopy; light-adapted states; photosynthetic reaction centers; protein conformational changes; rapid-scan Fourier transform infrared; two-dimensional correlation spectroscopy

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
1
Average
Average
Average
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