ENERGY TRANSFER IN TRIMERIC C-PHYCOCYANIN STUDIED BY PICOSECOND FLUORESCENCE KINETICS

The excited state kinetics of trimeric C-phycocyanin from Mastigocladus laminosus has been measured as a function of the emission and excitation wavelength by the single-photon timing technique with picosecond resolution and simultaneous data analysis. A fast decay component of 22 ps (C-phycocyanin with linker peptides) and 36 ps (C-phycocyanin lacking linker peptides) is attributed to efficient energy transfer from sensitizing to fluorescing chromophores. At long detection wavelengths the fast decay components are found to turn into a rise term. This finding further corroborates the concept of intramolecular energy transfer. Previous reports on the conformational heterogeneity of the chromophores and/or proteins in C-phycocyanin are confirmed. Our data also provide indications for the importance of the uncoloured linker peptides for this heterogeneity

The Effects of Alewife on the Zooplankton Community in Townhouse Pond

The purpose of this study was to determine what plankton species was more dominant in Townhouse Pond and what effects predation has on the food web. Plankton size is important when determining pelagic food web structure. Plankton size along with predation determines whether the lake is dominated by top-down or bottom-up control. Plankton size and biomass, grazing rates, light intensity, and chemistry parameters were all determined for Townhouse Pond in October 2008. We concluded that Townhouse Pond is a mesotrophic lake and is mainly dominated by phytoplankton, such as Microystis and Dinobryon. Alewife (Alosa pseudoharengus), planktivorous fish, are dominant in this pond resulting in top-down control. Although various studies were conducted here, further research could determine the abundance of alewife and their prey and specific parameter that effect their predation

Picosecond time-resolved energy transfer within C-phycocyanin aggregates of Mastigocladus laminosus

We have investigated by picosecond absorption experiments how the size of C-phycocyanin aggregates from Mastigocladus laminosus influences the excitation energy transfer kinetics. Going from C-phycocyanin monomers to trimers the lifetime of the faster energy transfer component decreased from 57 ± 4 to 27 ± 4 ps over most of the wavelength range (580–645 nm) studied. This change was interpreted as the opening of fast transfer channels (α-84 → β-84 and/or β-84 → β-84) between two adjacent monomers in the trimeric unit. The 57 ps lifetime is probably due mainly to the β-155 → β-84 energy transfer step. The intermediate lifetime decreased from about 300 ps in the monomer to 100–120 ps in the trimer. The former is believed to be dominated by the equilibration process α-84 a3 β-84, while the latter probably represents the time required for the excitation energy to reach thermodynamic equilibrium within the trimer. The lifetime of the longest components was about 1 ns in both systems. This indicates that the chromophores in these C-phycocyanin complexes are more exposed to non-radiative processes (like, for instance, isomerization) compared to the chromophores in intact phycobilisomes, where this lifetime typically is about 1.8 ns. The anisotropy relaxation closely followed the isotropic lifetimes in both systems. The anisotropy after the initial fast relaxation, r(∞), was 0.29 ± 0.04 in monomers and decreased to 0.15 ± 0.03 in trimers. Measurements of the steady-state fluorescence excitation anisotropy gave the same results within the experimental error

FÖRSTER TRANSFER CALCULATIONS BASED ON CRYSTAL STRUCTURE DATA FROM Agmenellum quadruplicatum C-PHYCOCYANIN

Excitation energy transfer in C-phycocyanin is modeled using the Forster inductive resonance mechanism. Detailed calculations are carried out using coordinates and orientations of the chromophores derived from X-ray crystallographic studies of C-phycocyanin from two different species (Schirmer et al, J. Mol. Biol. 184, 257–277 (1985) and ibid., 188, 651-677 (1986)). Spectral overlap integrals are estimated from absorption and fluorescence spectra of C-phycocyanin of Mastigocladus laminosus and its separated subunits. Calculations are carried out for the β-subunit, αβ-monomer, (αβ)3-trimer and (αβ)0-hexamer species with the following chromophore assignments: β155 = 's’(sensitizer), β84 =‘f (fluorescer) and α84 =‘m’(intermediate):]:. The calculations show that excitation transfer relaxation occurs to 3=98% within 200 ps in nearly every case; however, the rates increase as much as 10-fold for the higher aggregates. Comparison with experimental data on fluorescence decay and depolarization kinetics from the literature shows qualitative agreement with these calculations. We conclude that Forster transfer is sufficient to account for all of the observed fluorescence properties of C-phycocyanin in aggregation states up to the hexamer and in the absence of linker polypeptides

Picosecond time-resolved fluorescence of phycobiliproteins

The α- and β-subunits of C-phycocyanin from Mastigocladus laminosus were prepared according to revised procedures. Both subunits are isolated as dimers, which can be dissociated into monomers with detergent mixtures. The fluorescence decay kinetics are similar for the respective monomers and dimers. In no case could they be fitted by only one (α-subunit) or two exponentials (β-subunit) which are predicted by theory for samples with a unique chromophore—protein arrangement containing one and two chromophores, respectively. It is suggested that there exists a heterogeneity among the chromophores of the subunits, which may persist in the highly aggregated complexes present in cyanobacterial antennas

Assessment of Pelagic Food Webs in Mendums Pond, NH

This study focused on the relationship between plankton in Mendums Pond, NH. A grazing experiment was conducted to determine the effect of zooplankton on the phytoplankton population. The phytoplankton were largely composed of net plankton (75 %) and this fraction was dominated by cyanobacteria (84.5 %) even though this was a slightly acidic system. Data indicated that the mean body length of zooplankton increased with depth. The average body length of Daphnia ranged from 1.4 mm in the epilimnion to 1.9 mm in the hypolimnion. Copepods followed a similar trend increasing in average body length from 0.85 mm to 0.95 mm. The high numbers of cyanobacteria and copepods resulted in a 17.92 % day-1 grazing rate indicating that almost 18 % of the total lake water was filtered every day by the zooplankton. This also suggests that the phytoplankton are reproducing at a higher rate than they are being consumed by grazers. This may raise concerns about the future diversity of the food web as cyanobacteria reproduce and become more dominant in this system

BILIPROTEINS FROM THE BUTTERFLY Pieris brassicae STUDIED BY TIME-RESOLVED FLUORESCENCE AND COHERENT ANTI-STOKES RAMAN SPECTROSCOPY

The fluorescence decay time of the biliverdin IX7 chromophore present in biliproteins isolated from Pieris brassicae is determined to be 44 ± 3 ps. This value suggests a cyclic helical chromophore structure. The vibrational frequencies determined by CARS-spectroscopy are compared with those of model compounds. The data confirm that the chromophore in the protein-bound state adopts a cyclic-helical, flexible conformation

Fast preparative isoelectric focusing of phycocyanin subunits in layers of granulated gels

A new method is presented for the fast preparative separation of the light-harvesting photosynthetic pigment C-phycocanin into its and subunits, which is based on isoelectric focusing in layers of granutaled gels containing 7 M urea. The method has been successful in cases where other separation procedures failed. The recovery of the separated chains of the light -sensitive biliprotein amounts to 70 � 10 % when the separation is carried out under light exclusion and in an argon atmosphere. A simple and inexpensive setup for work under an atmosphere of protective gas is described