Charge transfer states in phycobilisomes

Phycobilisomes (PBs) absorb light and supply downstream photosynthetic processes with excitation energy in many cyanobacteria and algae. In response to a sudden increase in light intensity, excess excitation energy is photoprotectively dissipated in PBs by means of the orange carotenoid protein (OCP)-related mechanism or via a light-activated intrinsic decay channel. Recently, we have identified that both mechanisms are associated with far-red emission states. Here, we investigate the far-red states involved with the light-induced intrinsic mechanism by exploring the energy landscape and electro-optical properties of the pigments in PBs. While Stark spectroscopy showed that the far-red states in PBs exhibit a strong charge-transfer (CT) character at cryogenic temperatures, single molecule spectroscopy revealed that CT states should also be present at room temperature. Owing to the strong environmental sensitivity of CT states, the knowledge gained from this study may contribute to the design of a new generation of fluorescence markers

Stark fluorescence spectroscopy on peridinin–chlorophyll–protein complex of dinoflagellate, Amphidinium carterae

Because of their peculiar but intriguing photophysical properties, peridinin–chlorophyll–protein complexes (PCPs), the peripheral light-harvesting antenna complexes of photosynthetic dinoflagellates have been unique targets of multidimensional theoretical and experimental investigations over the last few decades. The major light-harvesting chlorophyll a (Chl a) pigments of PCP are hypothesized to be spectroscopically heterogeneous. To study the spectral heterogeneity in terms of electrostatic parameters, we, in this study, implemented Stark fluorescence spectroscopy on PCP isolated from the dinoflagellate Amphidinium carterae. The comprehensive theoretical modeling of the Stark fluorescence spectrum with the help of the conventional Liptay formalism revealed the simultaneous presence of three emission bands in the fluorescence spectrum of PCP recorded upon excitation of peridinin. The three emission bands are found to possess different sets of electrostatic parameters with essentially increasing magnitude of charge-transfer character from the blue to redder ones. The different magnitudes of electrostatic parameters give good support to the earlier proposition that the spectral heterogeneity in PCP results from emissive Chl a clusters anchored at a different sites and domains within the protein network

Identification and characterization of multiple emissive species in aggregated minor antenna complexes

Aggregation induced conformational change of light harvesting antenna complexes is believed to constitute one of the pathways through which photosynthetic organisms can safely dissipate the surplus of energy while exposed to saturating light. In this study, Stark fluorescence (SF) spectroscopy is applied to minor antenna complexes (CP24, CP26 and CP29) both in their light-harvesting and energy-dissipating states to trace and characterize different species generated upon energy dissipation through aggregation (in-vitro) induced conformational change. SF spectroscopy could identify three spectral species in the dissipative state of CP24, two in CP26 and only one in CP29. The comprehensive analysis of the SF spectra yielded different sets of molecular parameters for the multiple spectral species identified in CP24 or CP26, indicating the involvement of different pigments in their formation. Interestingly, a species giving emission around the 730 nm spectral region is found to form in both CP24 and CP26 following transition to the energy dissipative state, but not in CP29. The SF analyses revealed that the far red species has exceptionally large charge transfer (CT) character in the excited state. Moreover, the far red species was found to be formed invariably in both Zeaxanthin (Z)- and Violaxathin (V)-enriched CP24 and CP26 antennas with identical CT character but with larger emission yield in Z-enriched ones. This suggests that the carotenoid Z is not directly involved but only confers an allosteric effect on the formation of the far red species. Similar far red species with remarkably large CT character were also observed in the dissipative state of the major light harvesting antenna (LHCII) of plants [Wahadoszamen et al. PCCP, 2012], the fucoxanthin-chlorophyll protein (FCP) of brown algae [Wahadoszamen et al. BBA, 2014] and cyanobacterial IsiA [Wahadoszamen et al. BBA, 2015], thus pointing to identical sites and pigments active in the formation of the far red quenching species in different organisms

Phycocyanin: One Complex, Two States, Two Functions

Solar energy captured by pigments embedded in light-harvesting complexes can be transferred to neighboring pigments, dissipated, or emitted as fluorescence. Only when it reaches a reaction center is the excitation energy stabilized in the form of a charge separation and converted into chemical energy. Well-directed and regulated energy transfer within the network of pigments is therefore of crucial importance for the success of the photosynthetic processes. Using single-molecule spectroscopy, we show that phycocyanin can dynamically switch between two spectrally distinct states originating from two different conformations. Unexpectedly, one of the two states has a red-shifted emission spectrum. This state is not involved in energy dissipation; instead, we propose that it is involved in direct energy transfer to photosystem I. Finally, our findings suggest that the function of linker proteins in phycobilisomes is to stabilize one state or the other, thus controlling the light-harvesting functions of phycocyanin

Laser Raman Spectroscopy with Different Excitation Sources and Extension to Surface Enhanced Raman Spectroscopy

A dispersive Raman spectrometer was used with three different excitation sources (Argon-ion, He-Ne, and Diode lasers operating at 514.5 nm, 633 nm, and 782 nm, resp.). The system was employed to a variety of Raman active compounds. Many of the compounds exhibit very strong fluorescence while being excited with a laser emitting at UV-VIS region, hereby imposing severe limitation to the detection efficiency of the particular Raman system. The Raman system with variable excitation laser sources provided us with a desired flexibility toward the suppression of unwanted fluorescence signal. With this Raman system, we could detect and specify the different vibrational modes of various hazardous organic compounds and some typical dyes (both fluorescent and nonfluorescent). We then compared those results with the ones reported in literature and found the deviation within the range of ±2 cm−1, which indicates reasonable accuracy and usability of the Raman system. Then, the surface enhancement technique of Raman spectrum was employed to the present system. To this end, we used chemically prepared colloidal suspension of silver nanoparticles as substrate and Rhodamine 6G as probe. We could observe significant enhancement of Raman signal from Rhodamine 6G using the colloidal solution of silver nanoparticles the average magnitude of which is estimated to be 103

Laser Raman Spectroscopy with Different Excitation Sources and Extension to Surface Enhanced Raman Spectroscopy

A dispersive Raman spectrometer was used with three different excitation sources (Argon-ion, He-Ne, and Diode lasers operating at 514.5 nm, 633 nm, and 782 nm, resp.). The system was employed to a variety of Raman active compounds. Many of the compounds exhibit very strong fluorescence while being excited with a laser emitting at UV-VIS region, hereby imposing severe limitation to the detection efficiency of the particular Raman system. The Raman system with variable excitation laser sources provided us with a desired flexibility toward the suppression of unwanted fluorescence signal. With this Raman system, we could detect and specify the different vibrational modes of various hazardous organic compounds and some typical dyes (both fluorescent and nonfluorescent). We then compared those results with the ones reported in literature and found the deviation within the range of ±2 cm −1 , which indicates reasonable accuracy and usability of the Raman system. Then, the surface enhancement technique of Raman spectrum was employed to the present system. To this end, we used chemically prepared colloidal suspension of silver nanoparticles as substrate and Rhodamine 6G as probe. We could observe significant enhancement of Raman signal from Rhodamine 6G using the colloidal solution of silver nanoparticles the average magnitude of which is estimated to be 10 3

Absence of far-red emission band in aggregated core antenna complexes

Reported herein is a Stark fluorescence spectroscopy study performed on photosystem II core antenna complexes CP43 and CP47 in their native and aggregated states. The systematic mathematical modeling of the Stark fluorescence spectra with the aid of conventional Liptay formalism revealed that induction of aggregation in both the core antenna complexes via detergent removal results in a single quenched species characterized by a remarkably broad and inhomogenously broadened emission lineshape peaking around 700 nm. The quenched species possesses a fairly large magnitude of charge-transfer character. From the analogy with the results from aggregated peripheral antenna complexes, the quenched species is thought to originate from the enhanced chlorophyll-chlorophyll interaction due to aggregation. However, in contrast, aggregation of both core antenna complexes did not produce a far-red emission band at ∼730 nm, which was identified in most of the aggregated peripheral antenna complexes. The 730-nm emission band of the aggregated peripheral antenna complexes was attributed to the enhanced chlorophyll-carotenoid (lutein1) interaction in the terminal emitter locus. Therefore, it is very likely that the no occurrence of the far-red band in the aggregated core antenna complexes is directly related to the absence of lutein1 in their structures. The absence of the far-red band also suggests the possibility that aggregation-induced conformational change of the core antenna complexes does not yield a chlorophyll-carotenoid interaction associated energy dissipation channel