The effect of some antiseptic drugs on the energy transfer in chromatophore photosynthetic membranes of purple non-sulfur bacteria Rhodobacter sphaeroides.

Chromatophores of purple non-sulfur bacteria (PNSB) are invaginations of the cytoplasmic membrane that contain a relatively simple system of light-harvesting protein-pigment complexes, a photosynthetic reaction center (RC), a cytochrome complex, and ATP synthase, which transform light energy into the energy of synthesized ATP. The high content of negatively charged phosphatidylglycerol (PG) and cardiolipin (CL) in PNSB chromatophore membranes makes these structures potential targets that bind cationic antiseptics. We used the methods of stationary and kinetic fluorescence spectroscopy to study the effect of some cationic antiseptics (chlorhexidine, picloxydine, miramistin, and octenidine at concentrations up to 100 µM) on the spectral and kinetic characteristics of the components of the photosynthetic apparatus of Rhodobacter sphaeroides chromatophores. Here we present the experimental data on the reduced efficiency of light energy conversion in the chromatophore membranes isolated from the photosynthetic bacterium Rb. sphaeroides in the presence of cationic antiseptics. The addition of antiseptics did not affect the energy transfer between the light-harvesting LH1 complex and reaction center (RC). However, it significantly reduced the efficiency of the interaction between the LH2 and LH1 complexes. The effect was maximal with 100 µM octenidine. It has been proved that molecules of cationic antiseptics, which apparently bind to the heads of negatively charged cardiolipin molecules located in the rings of light-harvesting pigments on the cytoplasmic surface of the chromatophores, can disturb the optimal conditions for efficient energy migration in chromatophore membranes.

The mitochondrial-targeted reactive species scavenger JP4-039 prevents sulfite-induced alterations in antioxidant defenses, energy transfer, and cell death signaling in striatum of rats.

Sulfite oxidase (SO) deficiency is a disorder caused either by isolated deficiency of SO or by defects in the synthesis of its molybdenum cofactor. It is characterized biochemically by tissue sulfite accumulation. Patients present with seizures, progressive neurological damage, and basal ganglia abnormalities, the pathogenesis of which is not fully established. Treatment is supportive and largely ineffective. To address the pathophysiology of sulfite toxicity, we examined the effects of intrastriatal administration of sulfite in rats on antioxidant defenses, energy transfer, and mitogen-activated protein kinases (MAPK) and apoptosis pathways in rat striatum. Sulfite administration decreased glutathione (GSH) concentration and glutathione peroxidase, glucose-6-phosphate dehydrogenase, glutathione S-transferase, and glutathione reductase activities in striatal tissue. Creatine kinase (CK) activity, a crucial enzyme for cell energy transfer, was also decreased by sulfite. Superoxide dismutase-1 (SOD1) and catalase (CAT) proteins were increased, while heme oxygenase-1 (HO-1) was decreased. Additionally, sulfite altered phosphorylation of MAPK by decreasing of p38 and increasing of ERK. Sulfite further augmented the content of GSK-3ß, Bok, and cleaved caspase-3, indicating increased apoptosis. JP4-039 is a mitochondrial-targeted antioxidant that reaches higher intramitochondrial levels than other traditional antioxidants. Intraperitoneal injection of JP4-039 before sulfite administration preserved activity of antioxidant enzymes and CK. It also prevented or attenuated alterations in SOD1, CAT, and HO-1 protein content, as well as changes in p38, ERK, and apoptosis markers. In sum, oxidative stress and apoptosis induced by sulfite injection are prevented by JP4-039, identifying this molecule as a promising candidate for pharmacological treatment of SO-deficient patients.

Spectroscopic Investigation of DCCH and FTSC as a potential pair for Förster Resonance Energy Transfer in different solvents.

Two molecules, 7-(diethylamino)coumarin-3-carbohydrazide (DCCH) and fluorescein-5-thiosemicarbazide (FTSC) were investigated in different solvents, under varying pH conditions regarding their spectroscopic properties for the usage as a Förster Resonance Energy Transfer (FRET) pair to study the molecular interaction between cellulosic surfaces. All the relevant spectroscopic properties to determine the Förster distance were measured and the performance as a FRET system was checked. From the results, it is clear that the environmental conditions need to be accurately controlled as both, but especially the FTSC dyes are sensitive to changes. For high enough concentrations positive FRET systems were observed in DMF, DMSO, H2O, THF and alkaline DMF. However due to the low quantum yield of the unmodified DCCH throughout the investigated parameter range and the strong environmental dependency of FTSC, both dyes are not preferable for being used in a FRET system for studying interaction between cellulosic surfaces.

In-sights into the effect of heavy metal stress on the endogenous mustard cystatin.

Phytocystatins have been ascribed several protective roles against abiotic and biotic stress conditions. It was, therefore, thought worthwhile to document the effect of heavy metal stress on the endogenous plant cystatin. The mustard cystatin, purified from Brassica juncea (B. juncea) seeds retained its functional property of cysteine proteinase inhibition, despite exposure to high concentrations of metal ions, Cd2+ and Ni2+. An increase in inhibitory activity, ∼26% for Ni2+ and ∼16% for Cd2+ was observed, suggesting changes in protein conformation upon metal ion interaction. Isothermal calorimetric (ITC) studies show formation of a 1:1 binary complex on interaction with both metal ions but suggest a higher affinity for Ni2+. Fluorescence quenching data suggest a static quenching mechanism of interaction. Various spectroscopic analyses, namely, synchronous fluorescence, ANS fluorescence, far UV CD and ATR-FTIR spectroscopy show that the native mustard cystatin acquires a more ordered conformation upon interaction with metal ions. Differential Scanning Calorimetry indicates that the thermo-stability of the Ni2+ bound protein (Tm=109.4°C) is greater than both, the Cd2+ bound (Tm=104.5°C) and the native (Tm=99.5°C) forms. The B. juncea seed cystatin, is thus, identified as a potent and resilient member of the phytocystatin family with considerable inhibitory capacity despite exposure to heavy metal stress.

Chemical Denaturants Smoothen Ruggedness on the Free Energy Landscape of Protein Folding.

To characterize experimentally the ruggedness of the free energy landscape of protein folding is challenging, because the distributed small free energy barriers are usually dominated by one, or a few, large activation free energy barriers. This study delineates changes in the roughness of the free energy landscape by making use of the observation that a decrease in ruggedness is accompanied invariably by an increase in folding cooperativity. Hydrogen exchange (HX) coupled to mass spectrometry was used to detect transient sampling of local energy minima and the global unfolded state on the free energy landscape of the small protein single-chain monellin. Under native conditions, local noncooperative openings result in interconversions between Boltzmann-distributed intermediate states, populated on an extremely rugged "uphill" energy landscape. The cooperativity of these interconversions was increased by selectively destabilizing the native state via mutations, and further by the addition of a chemical denaturant. The perturbation of stability alone resulted in seven backbone amide sites exchanging cooperatively. The size of the cooperatively exchanging and/or unfolding unit did not depend on the extent of protein destabilization. Only upon the addition of a denaturant to a destabilized mutant variant did seven additional backbone amide sites exchange cooperatively. Segmentwise analysis of the HX kinetics of the mutant variants further confirmed that the observed increase in cooperativity was due to the smoothing of the ruggedness of the free energy landscape of folding of the protein by the chemical denaturant.

Disruption of Energy Transfer and Redox Status by Sulfite in Hippocampus, Striatum, and Cerebellum of Developing Rats.

Patients with sulfite oxidase (SO) deficiency present severe brain abnormalities, whose pathophysiology is not yet elucidated. We evaluated the effects of sulfite and thiosulfate, metabolites accumulated in SO deficiency, on creatine kinase (CK) activity, mitochondrial respiration and redox status in hippocampus, striatum and cerebellum of developing rats. Our in vitro results showed that sulfite and thiosulfate decreased CK activity, whereas sulfite also increased malondialdehyde (MDA) levels in all brain structures evaluated. Sulfite further diminished mitochondrial respiration and increased DCFH oxidation and hydrogen peroxide production in hippocampus. Sulfite-induced CK activity decrease was prevented by melatonin (MEL), resveratrol (RSV), and dithiothreitol while increase of MDA levels was prevented by MEL and RSV. Regarding the antioxidant system, sulfite increased glutathione concentrations in hippocampus and striatum. In addition, sulfite decreased the activities of glutathione peroxidase in all brain structures, of glutathione S-transferase in hippocampus and cerebellum, and of glutathione reductase in cerebellum. In vivo experiments performed with intrahippocampal administration of sulfite demonstrated that this metabolite increased superoxide dismutase activity without altering other biochemical parameters in rat hippocampus. Our data suggest that impairment of energy metabolism and redox status may be important pathomechanisms involved in brain damage observed in individuals with SO deficiency.

Incorporation of spheroidene and spheroidenone into light-harvesting complexes from purple sulfur bacteria.

Spheroidene and spheroidenone from the non-sulfur bacterium Rhodobacter (Rba.) sphaeroides were incorporated into diphenylamine (DPA) LH1-RC and LH2 complexes from sulfur bacteria Allochromatium (Alc.) minutissimum and Ectothiorhodospira (Ect.) haloalkaliphila in which carotenoid (Car) biosynthesis was inhibited by ~95%. A series of biochemical characteristics of the modified LH2 complexes was studied (electrophoretic mobility, absorption and CD spectra, Car composition, Car-to-BChl energy transfer and thermal stability). It was found that the electrophoretic mobility of the complexes with incorporated Cars did not change compared to that of the control and DPA-complexes, indicating the absence of any significant change in the structure of LH complexes upon DPA-treatment and subsequent incorporation of Cars. The analysis of fluorescence excitation spectra of the spheroidene-incorporated LH2 complex (LH2:sph) and the spheroidenone-incorporated LH2 complex (LH2:sph-ne) showed that spheroidene and spheroidenone exhibited relatively low efficiencies of energy transfer to BChl, when incorporated into the LH2 DPA-complexes from Alc. minutissimum and Ect. haloalkaliphila, although, they showed high efficiencies, being in their natural state in the LH2 complexes from Rba. sphaeroides. A significant increase in thermostability observed for the LH2:sph and LH2:sph-ne complexes with respect to the LH2 DPA-complexes indicated that the two incorporated Cars stabilized the structure of the LH2 complexes.

Fluorescent Property of Chitosan Oligomer and Its Application as a Metal Ion Sensor.

An aqueous solution was successfully prepared using a low-molecular-weight chitosan oligomer and FITC, and its structural and fluorescent properties were observed by using ¹H NMR, 13C NMR, FT-IR, XRD, UV-Vis, and PL spectrometry. Its application as a metal ion sensor was also evaluated. The fluorescence in the water-soluble chitosan oligomer was a result of the carbamato anion (NHCOO-), and a synthesized FITC-labeled chitosan oligomer exhibited an effective detection effect for copper ion as well as energy transfer by the ion near FITC that caused a fluorescence decrease (quenching). The chitosan oligomer was confirmed to be applicable as a selective and sensitive colorimetric sensor to detect Cu2+.

Mapping of voltage sensor positions in resting and inactivated mammalian sodium channels by LRET.

Voltage-gated sodium channels (Navs) play crucial roles in excitable cells. Although vertebrate Nav function has been extensively studied, the detailed structural basis for voltage-dependent gating mechanisms remain obscure. We have assessed the structural changes of the Nav voltage sensor domain using lanthanide-based resonance energy transfer (LRET) between the rat skeletal muscle voltage-gated sodium channel (Nav1.4) and fluorescently labeled Nav1.4-targeting toxins. We generated donor constructs with genetically encoded lanthanide-binding tags (LBTs) inserted at the extracellular end of the S4 segment of each domain (with a single LBT per construct). Three different Bodipy-labeled, Nav1.4-targeting toxins were synthesized as acceptors: ß-scorpion toxin (Ts1)-Bodipy, KIIIA-Bodipy, and GIIIA-Bodipy analogs. Functional Nav-LBT channels expressed in Xenopus oocytes were voltage-clamped, and distinct LRET signals were obtained in the resting and slow inactivated states. Intramolecular distances computed from the LRET signals define a geometrical map of Nav1.4 with the bound toxins, and reveal voltage-dependent structural changes related to channel gating.

Viscous and fermentable nonstarch polysaccharides affect intestinal nutrient and energy flow and hindgut fermentation in growing pigs.

Two experiments were conducted to determine the effects of viscosity and fermentability of purified nonstarch polysaccharides (NSP) on intestinal nutrient and energy flow and hindgut fermentation in growing pigs. In Exp. 1, 15 ileal-cannulated pigs (50.5 ± 2.9 kg BW) were allotted to 5 diets in a 2-period incompletely randomized design. Pigs were provided a cornstarch N-free diet (CST) or a diet containing 5% inulin (INU), carboxymethylcellulose sodium (CMC), microcrystalline cellulose (MCC), or Solka-Floc (SFC). Apparent ileal digestibility (AID) of DM, ash, and GE were greater ( < 0.05) in the MCC and SFC diets than in the INU and CMC diets. Apparent total tract digestibility (ATTD) of DM, carbohydrates (CHO), and GE and the DE content in the CST and INU diets were greater ( < 0.01) than in the CMC, MCC, and SFC diets. Hindgut disappearance of DM, CHO, and GE in the INU diet was greater ( < 0.05) than in the other N-free diets. The ileal endogenous flow of His, Ile, Leu, Met, Thr, Val, and all dispensable AA in the CMC diet was greater ( < 0.05) than in the other diets. In Exp. 2, 6 ileal-cannulated pigs were allotted to 3 diets containing either a corn-soybean meal control diet or the control diet with 5% INU or CMC in a replicated 3 × 3 Latin square design. Standardized ileal digestibility (SID) of Ala, Arg, Asp, Cys, Gly, His, Ile, Leu, Lys, Phe, Ser, Thr, and Val were greater ( < 0.05) in the CMC diet than in the control and INU diets. Incorporation of 5% INU or CMC in the control diet reduced ( < 0.01) the AID of arabinoxylan and insoluble and total NSP. The ATTD of NSP and cellulose and the hindgut disappearance of NSP, ß-glucan, and cellulose in the CMC diet were less ( < 0.01) than in the control and INU diets. Inclusion of 5% INU in the diet increased ( < 0.01) hindgut fermentation of insoluble and total NSP compared with the control and CMC diets. In conclusion, depending on the viscosity and fermentability of the NSP, different sources will have different effects on nutrient digestibility and hindgut fermentation. Addition of 5% INU to a corn-soybean meal diet reduced digestibility of the NSP component in the ileum and increased hindgut fermentation of total NSP. In contrast, the inclusion of CMC increased the AID and SID of the diet and reduced total tract digestion and hindgut fermentation of NSP component. Carboxymethylcellulose sodium is not recommended as a source of synthetic fiber in a N-free diet to determine the SID of AA of diets.

Caffeine ingestion improves power output decrement during 3-min all-out exercise.

PURPOSE: To investigate the effect of caffeine ingestion on the 3-min all-out test (3MT) performance and plasma electrolytes in athletes. METHODS: Fifteen collegiate male basketball players were recruited and completed two trials separated by at least 1 week in caffeine (CAF, 6 mg kg(-1)) and placebo conditions. During the first visit, participants performed an incremental cycling test to determine their 3MT resistance. After a familiarization trial, participants performed a CAF or PL trial according to a randomized crossover design. One hour after ingesting capsules, the participants performed the 3MT to estimate the end-test power (EP) and work done above EP (WEP). Blood samples for sodium (Na(+)), potassium (K(+)), pH, and lactate concentrations were drawn pretest, 1 h after ingestion, and posttest. RESULTS: Significant differences in WEP (CAF vs. PL, 13.4 ± 3.0 vs. 12.1 ± 2.7 kJ, P < 0.05) but not in EP (CAF vs. PL, 242 ± 37 vs. 244 ± 42 W, P > 0.05) were determined between the conditions. Compared with the PL condition, the CAF condition yielded significantly higher power outputs (60-150 s), a lower fatigue rate during the 3MT (CAF vs. PL, 0.024 ± 0.007 vs. 0.029 ± 0.006 s(-1), P < 0.05), a significantly higher lactate concentration after the 3MT, and significantly lower K(+) concentrations at 1 h after caffeine ingestion. There were no significant interaction effects for pH and Na(+) concentrations. CONCLUSIONS: Caffeine ingestion did not change EP but improved WEP and the rate of decline in power output during short-term, severe exercise.

Differences in energy transfer of a cyanobacterium, Synechococcus sp. PCC 7002, grown in different cultivation media.

Currently, cyanobacteria are regarded as potential biofuel sources. Large-scale cultivation of cyanobacteria in seawater is of particular interest because seawater is a low-cost medium. In the present study, we examined differences in light-harvesting and energy transfer processes in the cyanobacterium Synechococcus sp. PCC 7002 grown in different cultivation media, namely modified A medium (the optimal growth medium for Synechococcus sp. PCC 7002) and f/2 (a seawater medium). The concentrations of nitrate and phosphate ions were varied in both media. Higher nitrate ion and/or phosphate ion concentrations yielded high relative content of phycobilisome. The cultivation medium influenced the energy transfers within phycobilisome, from phycobilisome to photosystems, within photosystem II, and from photosystem II to photosystem I. We suggest that the medium also affects charge recombination at the photosystem II reaction center and formation of a chlorophyll-containing complex.

Porphyra-334 isolated from the marine algae Bangia atropurpurea: conformational performance for energy conversion.

Prophyra-334 (p-334) may play a role of energy transfer under an uncertain mechanism, and we speculate the possible model. Via 1D and 2D NMR experiments, it was simulated the correlation between dissociation and conformation of p-334. Intramolecular interactions were observed based on a series of changes in the 1H and 13C chemical shifts. Nuclear Overhauser effect spectroscopy experiments and molecular models in various pD conditions indicated the p-334 molecular dissociation process status. In addition, we also used Chem3D software to find the most possible molecular conformation. The relationship between the structural status and energy conversion is explained. Those are the primary results. More researches on it are highly expected in the future.

Roles of the cyclic electron flow around PSI (CEF-PSI) and O2-dependent alternative pathways in regulation of the photosynthetic electron flow in short-term fluctuating light in Arabidopsis thaliana.

To assess the roles of the cyclic electron flow around PSI (CEF-PSI) and O2-dependent alternative pathways including the water-water cycle in fluctuating light, we grew the wild type and pgr5 mutant of Arabidopsis thaliana in constant light, and measured Chl fluorescence and P700 parameters in their leaves in the fluctuating light alternating between 240 (HL) and 30 µmol photons m⁻² s⁻² (LL) every 2 min. At 20% O2, the photochemical quantum yield of PSII decreased, in particular in the pgr5 plants, soon after the start of the fluctuating light treatment. PSI of the pgr5 plants was markedly photoinhibited by this treatment for 42 min. Slight PSI photoinhibition was also observed in the wild type. We measured energy sharing between PSII and PSI and estimated the PSI and PSII electron transport rates (ETRs). pgr5 showed larger energy allocation to PSI. In contrast to the wild type, the ratio of the PSI to the PSII ETR in pgr5 was higher in LL but lower in HL at 20% O2 due to PSI acceptor-side limitation. At 2.7% or 0% O2, the CEF-PSI of the pgr5 plants was enhanced, the acceptor-side limitation of PSI electron flow was released and PSI photoinhibition was not observed. The results suggest that the light fluctuation is a potent stress to PSI and that the CEF-PSI is essential to protect PSI from this stress.

Insights into the selective binding and toxic mechanism of microcystin to catalase.

Microcystin is a sort of cyclic nonribosomal peptides produced by cyanobacteria. It is cyanotoxin, which can be very toxic for plants and animals including humans. The present study evaluated the interaction of microcystin and catalase, under physiological conditions by means of fluorescence, three-dimensional (3D) fluorescence, circular dichroism (CD), Fourier Transform infrared (FT-IR) spectroscopy, and enzymatic reactionkinetic techniques. The fluorescence data showed that microcystin could bind to catalase to form a complex. The binding process was a spontaneous molecular interaction procedure, in which electrostatic interactions played a major role. Energy transfer and fluorescence studies proved the existence of a static binding process. Additionally, as shown by the three-dimensional fluorescence, CD and FT-IR results, microcystin could lead to conformational and microenvironmental changes of the protein, which may affect the physiological functions of catalase. The work provides important insights into the toxicity mechanism of microcystin in vivo.

A comparative study on the binding of single and double chain surfactant-cobalt(III) complexes with bovine serum albumin.

The comparative binding effect of single and double aliphatic chain containing surfactant-cobalt(III) complexes cis-[Co(bpy)2(DA)2](ClO4)3·2H2O (1), cis-[Co(bpy)2(DA)Cl](ClO4)2·2H2O (2), cis-[Co(phen)2(CA)2](ClO4)3·2H2O (3), and cis-[Co(phen)2(CA)Cl](ClO4)2·2H2O (4) with bovine serum albumin (BSA) under physiological condition was analyzed by steady state, time resolved fluorescence, synchronous, three-dimensional fluorescence, UV-Visible absorption and circular dichroism spectroscopic techniques. The results show that these complexes cause the fluorescence quenching of BSA through a static mechanism. The binding constants (Kb) and the number of binding sites were calculated and binding constant values are found in the range of 10(4)-10(5) M(-1). The results indicate that compared to single chain complex, double chain surfactant-cobalt(III) complex interacts strongly with BSA. Also the sign of thermodynamic parameters (ΔG°, ΔH°, and ΔS°) indicate that all the complexes interact with BSA through hydrophobic force. The binding distance (r) between complexes and BSA was calculated using Förster non-radiation energy transfer theory and found to be less than 7 nm. The results of synchronous, three dimensional fluorescence and circular dichroism spectroscopic methods indicate that the double chain surfactant-cobalt(III) complexes changed the conformation of the protein considerably than the respective single chain surfactant-cobalt(III) complexes. Antimicrobial studies of the complexes showed good activities against pathogenic microorganisms.

Spectroscopic studies on the interaction of carteolol hydrochloride and urea-induced bovine serum albumin.

The interaction of carteolol hydrochloride, to 0.2 mol L(-1) urea-induced bovine serum albumin in aqueous solution has been first investigated by fluorescence spectra and ultraviolet-visible (UV-vis) spectra at pH 7.40. The quenching mechanism, binding parameter and sites (n), the binding mode (ΔG, ΔH, and ΔS) as well as the binding distance (r) have been obtained according to the experimental results. We also use the synchronous fluorescence method to study the effect of CTL on the conformation change of urea-induced BSA.

Energy-dissipative supercomplex of photosystem II associated with LHCSR3 in Chlamydomonas reinhardtii.

Plants and green algae have a low pH-inducible mechanism in photosystem II (PSII) that dissipates excess light energy, measured as the nonphotochemical quenching of chlorophyll fluorescence (qE). Recently, nonphotochemical quenching 4 (npq4), a mutant strain of the green alga Chlamydomonas reinhardtii that is qE-deficient and lacks the light-harvesting complex stress-related protein 3 (LHCSR3), was reported [Peers G, et al. (2009) Nature 462(7272):518-521]. Here, applying a newly established procedure, we isolated the PSII supercomplex and its associated light-harvesting proteins from both WT C. reinhardtii and the npq4 mutant grown in either low light (LL) or high light (HL). LHCSR3 was present in the PSII supercomplex from the HL-grown WT, but not in the supercomplex from the LL-grown WT or mutant. The purified PSII supercomplex containing LHCSR3 exhibited a normal fluorescence lifetime at a neutral pH (7.5) by single-photon counting analysis, but a significantly shorter lifetime at pH 5.5, which mimics the acidified lumen of the thylakoid membranes in HL-exposed chloroplasts. The switch from light-harvesting mode to energy-dissipating mode observed in the LHCSR3-containing PSII supercomplex was sensitive to dicyclohexylcarbodiimide, a protein-modifying agent specific to protonatable amino acid residues. We conclude that the PSII-LHCII-LHCSR3 supercomplex formed in the HL-grown C. reinhardtii cells is capable of energy dissipation on protonation of LHCSR3.

Conjugated polymers for light-activated antifungal activity.

A cationic polythiophene-porphyrin (PTP) dyad is shown to exhibit efficient light-activated antifungal activity. Higher singlet oxygen (¹O2) generation efficiency can be attained from PTP upon photoexcitation due to the light-harvesting properties of the polymer backbone and efficient energy transfer from the polythiophene to the porphyrin units. PTP can be used for treating fungal infections in lower doses of irradiation light and polymer concentration.

Stabilization and modulation of the phycobilisome by calcium in the calciphilic freshwater red alga Bangia atropurpurea.

The bangiophycean filamentous red alga Bangia atropurpurea is distributed in freshwater habitats such as littoral and splash zones of lakes or rapid currents distant from the sea. In these habitats, the distribution and growth of this alga appear to be related to hard water rich in calcium ions. To characterize the eco-physiological properties of this calciphilic red alga, we examined the effects of long-term and short-term Ca(2+) depletion on photosynthetic growth of the thallus and on the phycobilisome. Long-term culture experiments suggested that higher Ca(2+) concentrations (>50mgL(-1)) were required to sustain thallus growth and pigmentation of cells. In short-term Ca(2+)-depletion treatments, fluorescence derived from phycoerythrin (PE) fluctuated, although the absorption spectra of the thalli did not change. After 30 min of Ca(2+) depletion, the fluorescence lifetime of PE became markedly longer, indicating that the energy transfer from PE to phycocyanin (PC) was suppressed. The fluorescence lifetime of PE returned to its original value within a short time after 4h of Ca(2+) depletion, however, energy transfer from PE to PC was still suppressed. This suggested that the excitation energy absorbed by PE was quenched during prolonged Ca(2+) depletion. The efficient energy transfer from PC and allophycocyanin were unchanged during these treatments.