Photophysics of phycoerythrocyanins from the cyanobacterium Westiellopsis prolifica studied by time-resolved fluorescence and coherent anti-Stokes Raman scattering spectroscopy

Three building blocks of the antenna complexes of the cyanobacterium Westiellopsis prolifica were studied: PEC(X), which is similar to the α-subunit of phycoerythrocyanin (PEC), trimers of PEC and monomers derived from these by deaggregation with KSCN. The fit of the fluorescence decay curve of PEC(X) requires at least four exponentials, although it supposedly contains only one chromophore. The coherent anti-Stokes Raman scattering (CARS) spectra indicate that the heterogeneity observed is due to geometrical isomers, which are in part generated by photoinduced processes. A similar heterogeneity in chromophore structure and properties is also found in the monomers, where four exponentials are needed to fit the fluorescence decay curve. As in trimers, there is a long-lived, low-amplitude component, which can be assigned to impurities and/or oxidation products. The energy transfer time between the two phyocyanobilin chromophores in the β-subunit is about 500 ps; the lifetime of the fluorescing β-chromophore is 1.5 ns. The phycoviolobilin chromophore in the α-subunit adopts different geometries characterized by fluorescence lifetimes of about 240 and 800 ps. No evidence was found for energy transfer between the α-chromophore and the β-chromophores. This energy transfer occurs in trimers on a time scale of less than 20 ps; the energy transfer time between the two different types of β-chromophore is about 250 ps and the lifetime of the terminal emitter is about 1.5 ns. The excited state kinetics are therefore similar to those of PEC trimers from Mastigocladus laminosus, as are the CARS spectra, indicating a similar chromophore—protein arrangement. In comparison with phycocyanin, the ordering of the excited states of chromophores β84 and β155 may be changed. Although PEC trimers of Westiellopsis prolifica show almost as good a photostability as trimers of Mastigocladus laminosus, monomers are so photolabile that no CARS spectra could be recorded

FLUORESCENCE AND CIRCULAR DICHROISM STUDIES ON THE PHYCOERYTHROCYANINS FROM THE CYANOBACTERIUM

Two phycoerythrocyanin (PEC) fractions have been obtained from the phycobilisomes of the cyanobac-terium Westiellopsis prolifica ARM 365. They have been characterized by absorption, fluorescence and circular dichroism spectroscopy. One of them is spectroscopically similar to a PEC trimer known from other organisms. Whereas efficient energy transfer from its violin (α-84) to the cyanin (β-84, 155) chromophores is efficient in the trimer (αβ it is impeded after dissociation to the monomer (α,β). A second fraction of PEC which we earlier termed PEC(X) (Maruthi Sai et al., Photochem. Photobiol. 55,119–124, 1992), exhibited the spectral properties similar to that of the α-subunit of PEC from Mastigocladus laminosus. With this highly photoactive fraction, the circular dichroism spectra of the violobilin chromophore in both photoreversible states were obtained

Förster energy transfer between neighbouring chromophores in C-phycocyanin trimers

The excitation-energy transfer in C-phycocyanin (C-PC) trimers and monomers isolated from phycobilisomes of Mastigocladus laminosus has been studied by polarization femtosecond laser spectroscopy. Excitation with 70-fs pulses at 615 nm gave rise to a 500-fs energy-transfer process that was observed only in trimeric preparations. The rate of the process is in agreement with earlier calculated Förster energy transfer rates between neighbouring α-84 and β-84 chromophores of different monomeric subunits. This process is most clearly seen in the anisotropy decay kinetics. As a result of femtosecond excitation-energy transfer, the anisotropy relaxes from 0.4 to 0.23. The final anisotropy value is in fair agreement with the results of calculations based on the crystal structure and spectroscopic data of C-PC trimers. Our results support the conclusion that Förster energy transfer can occur between excitonically coupled chromophores

Equivalence between the real time Feynman histories and the quantum shutter approaches for the "passage time" in tunneling

We show the equivalence of the functions $G_{\rm p}(t)$ and $|\Psi(d,t)|^2$ for the ``passage time'' in tunneling. The former, obtained within the framework of the real time Feynman histories approach to the tunneling time problem, using the Gell-Mann and Hartle's decoherence functional, and the latter involving an exact analytical solution to the time-dependent Schr\"{o}dinger equation for cutoff initial waves

The role of insulin receptor substrate 2 in hypothalamic and β cell function

Insulin receptor substrate 2 (Irs2) plays complex roles in energy homeostasis. We generated mice lacking Irs2 in β cells and a population of hypothalamic neurons (RIPCreIrs2KO), in all neurons (NesCreIrs2KO), and in proopiomelanocortin neurons (POMCCreIrs2KO) to determine the role of Irs2 in the CNS and β cell. RIPCreIrs2KO mice displayed impaired glucose tolerance and reduced β cell mass. Overt diabetes did not ensue, because β cells escaping Cre-mediated recombination progressively populated islets. RIPCreIrs2KO and NesCreIrs2KO mice displayed hyperphagia, obesity, and increased body length, which suggests altered melanocortin action. POMCCreIrs2KO mice did not display this phenotype. RIPCreIrs2KO and NesCreIrs2KO mice retained leptin sensitivity, which suggests that CNS Irs2 pathways are not required for leptin action. NesCreIrs2KO and POMCCreIrs2KO mice did not display reduced β cell mass, but NesCreIrs2KO mice displayed mild abnormalities of glucose homeostasis. RIPCre neurons did not express POMC or neuropeptide Y. Insulin and a melanocortin agonist depolarized RIPCre neurons, whereas leptin was ineffective. Insulin hyperpolarized and leptin depolarized POMC neurons. Our findings demonstrate a critical role for IRS2 in β cell and hypothalamic function and provide insights into the role of RIPCre neurons, a distinct hypothalamic neuronal population, in growth and energy homeostasis

Influence of chromophores on quarternary structure of phycobiliproteins from the cyanobacterium, Mastigocladus laminosus

Chromophores of C-phycocyanin and phycoerythrο-cyanin have been chemically modified by reduction to rubins , bleaching , photoisomerization , or perturbation with bulky substituents. Pigments containing modified chromophores, or hybrids containing modified and unmodified chromophores in individual protomers have been prepared. All modifications inhibit the association of the (aß)-protomers of these pigments to higher aggregates. The results demonstrate a pronounced effect of the state of the chromophores on biliprotein quaternary structure. It may be important in phycobi1isome assembly , and also in the dual function of biliproteins as (i) antenna pigments for photosynthesis and (ii) reaction centers for photomor-phogenesis

The role of insulin receptor substrate 2 in hypothalamic and beta cell function

Insulin receptor substrate 2 (Irs2) plays complex roles in energy homeostasis. We generated mice lacking Irs2 in beta cells and a population of hypothalamic neurons (RIPCreIrs2KO), in all neurons (NesCreIrs2KO), and in proopiomelanocortin neurons (POMCCreIrs2KO) to determine the role of Irs2 in the CNS and beta cell. RIPCreIrs2KO mice displayed impaired glucose tolerance and reduced P cell mass. Overt diabetes did not ensue, because beta cells escaping Cre-mediated recombination progressively populated islets. RIPCreIrs2KO and NesCreIrs2KO mice displayed hyperphagia, obesity, and increased body length, which suggests altered melanocortin action. POMCCreIrs2KO mice did not display this phenotype. RIPCreIrs2KO and NesCreIrs2KO mice retained leptin sensitivity, which suggests that CNS Irs2 pathways are not required for leptin action. NesCreIrs2KO and POMCCreIrs2KO mice did not display reduced beta cell mass, but NesCreIrs2KO mice displayed mild abnormalities of glucose homeostasis. RIPCre neurons did not express POMC or neuropeptide Y. Insulin and a melanocortin agonist depolarized RIPCre neurons, whereas leptin was ineffective. Insulin hyperpolarized and leptin depolarized POMC neurons. Our findings demonstrate a critical role for IRS2 in beta cell and hypothalamic function and provide insights into the role of RIPCre neurons, a distinct hypothalamic neuronal population, in growth and energy homeostasis

Femtosecond spectral and anisotropy study of excitation energy transfer between neighbouring α-80 and β-81 chromophores of allophycocyanin trimers

Polarization pump-probe femtosecond spectroscopy was used to investigate photoinduced optical density changes in allophycocyanin (APC) trimers at 635–690 nm after excitation with 230-fs pulses at 618 nm. The initial bleaching observed at λ < 645 nm is followed by subpicosecond absorption recovery corresponding to 430 ± 40 fs recovery kinetics measured at 615 nm with 70-fs pulses. Only the red part of the APC absorption band remains strongly bleached at 3 ps after excitation. The spectral and kinetic results can be described in terms of two different models of interaction between neighbouring α-80 and β-81 chromophores of APC trimers. According to the first one, the observed subpicosecond kinetics corresponds to relaxation between the levels of excitonically coupled, spectrally identical α-80 and β-81 chromophores. Excited state absorption to doubly excited excitonic state should in this case contribute to the measured difference spectra. According to the second one, the femtosecond excitation energy transfer in APC trimers takes place between a donor chromophore absorbing predominantly at 620 nm and an acceptor chromophore absorbing at 650 nm. The high anisotropy value observed at 615 nm during the first 1.2 ps is in good agreement with the donor-acceptor model. Anisotropy values calculated in the 635–675 nm spectral region at 3 ps after excitation are in the 0.1–0.25 range corresponding to an angle of 30°–45° between donor and acceptor transition dipole orientations. The high anisotropy obtained at 658 nm during the excitation is probably due to stimulated emission of the donor chromophore

PHOTOCHEMISTRY OF PHYCOBILIPROTEINS

Native PEC from the cyanobacterium, Mastigocladus laminosus, and its isolated α-subunit show photoreversibly photochromic reactions with difference-maxima around 502 and 570 nm in the spectral region of the α-84 phycoviolobilin chromophore. (b) Native PEC and its β-subunit show little if any reversible photochemistry in the 600–620 nm region, where the phycocyanobilin chromophores on the β-subunit absorb maximally, (c) Reversible photochemistry is retained in ureadenatured PEC at pH = 7.0 or pH ≤ 3. The difference maxima are shifted to 510 and 600 nm, and the amplitudes are decreased. An irreversible absorbance increase occurs around 670 nm (pH ≤ 3). (d) The amplitude of the reversible photoreaction difference spectrum is maximum in the presence of 4–5 M urea or 1 M KSCN, conditions known to dissociate phycobiliprotein aggregates into monomers. At the same time, the phycocyanobilin chromophore(s) are bleached irreversibly, (e) The amplitude becomes very small in high aggregates, e.g. in phycobilisomes. (f) In a reciprocal manner, the phototransformation of native PEC leads to a reversible shift of its aggregation equilibrium between trimer and monomer. The latter is favored by orange, the former by green light, (g) It is concluded that the phycoviolobilin chromophore of PEC is responsible for reversible photochemistry in PEC, and that there is not only an influence of aggregation state on photochemistry, but also vice versa an effect of the status of the chromophore on aggregation state. This could constitute a primary signal in the putative function as sensory pigment, either directly, or indirectly via the release of other polypeptides, via photodynamic effects, or the like

Dual-polarized aperture-coupled patch antennas with application to retrodirective and monopulse arrays

An isolation technique, which does not require conventional circulators, is proposed for the realization of a simple and low-cost aperture-coupled circularly polarized antenna for application to full-duplex devices. The approach is based on the use of slotlines loops to provide surface current cancellation in specific regions of the antenna structure, leading to improved axial ratio and isolation between the ports in excess of 50 dB. Circular polarization is achieved by introducing a double-box hybrid coupler, which is optimized to obtain good matching and isolation of the quadrature signals. On this basis, both right- and left-hand circularly polarized beams are achieved by interchanging the transmitting and receiving antenna ports, enabling full-duplex operation and reconfigurability. While the antenna structure is designed for 2.45 GHz operation, one can take advantage of the proposed approach to tune the frequency of maximum isolation. Both single-element prototypes as well as a 2 × 2 array are fabricated and measured, showing good agreement with the simulations and validating the proposed isolation approach. The beam steering capabilities as well as the application to a Van Atta retrodirective antenna array and the possibilities of achieving delta and sum patterns for monopulse operation are also reported. The proposed full-duplex antenna can also represent an excellent solution for narrowband wireless power transmission systems