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At least 883 records · Page 49Linked to original sources
PubMed1990

Purification and Characterization of Glutamyl-tRNA Synthetase : An Enzyme Involved in Chlorophyll Biosynthesis.

Chlorophyll biosynthesis starts with the synthesis of glutamyl-tRNA (glu-tRNA) by a glutamyl-tRNA synthetase (Glu RS). The glu-tRNA is subsequently transformed to delta-aminolevulinic acid (ALA), which is a committed and regulated precursor in the chlorophyll biosynthetic pathway. The Glu RS from a green alga, Chlamydomonas reinhardtii, was purified and shown to be able to synthesize glu-tRNA and to participate in ALA synthesis in a coupled enzyme assay. Physical and chemical characterization of the purified Glu RS indicated that the enzyme had been purified to homogeneity. The purified enzyme has a native molecular weight of 60,000, an isoelectric point of 4.6, and it formed a single band of 32,500 daltons when analyzed by a silver stained denaturing gel. The N-terminal amino acid sequence of the 32,500 dalton protein was determined to be Asn-Lys-Val-Ala-Leu-Leu-Gly-Ala-Ala-Gly. The molecular weight analyses together with the unambiguous N-terminal amino acid sequence obtained from the purified enzyme suggested that the native enzyme was composed of two identical subunits. Polyclonal antibodies raised against the purified and denatured enzyme were able to inhibit the activity of the native enzyme and to interact specifically with the 32,500 dalton band on Western blots. Thus, the antibodies provided an additional linkage for the structural and functional identities of the enzyme. In vitro experiments showed that over 90% of the glu RS activity was inhibited by 5 micromolar heme, which suggested that Glu RS may be a regulated enzyme in the chlorophyll biosynthetic pathway.

Journal Article↗
PubMed2003

Validation of Terra-MODIS phytoplankton chlorophyll fluorescence line height. I. Initial airborne lidar results.

The Moderate Resolution Imaging Spectroradiometer (MODIS) aboard the Terra spacecraft contains spectral bands that allow retrieval of solar-induced phytoplankton chlorophyll fluorescence emission radiance. Concurrent airborne laser-induced (and water-Raman normalized) phytoplankton chlorophyll fluorescence data is used to successfully validate the MODIS chlorophyll fluorescence line height (FLH) retrievals within Gulf Stream, continental slope, shelf, and coastal waters of the Middle Atlantic Bight portion of the western North Atlantic Ocean for 11 March 2002. Over the entire approximately 480-km flight line a correlation coefficient of r2 = 0.85 results from regression of the airborne laser data against the MODIS FLH. It is also shown that the MODIS FLH product is not influenced by blue-absorbing chromophoric dissolved organic matter absorption. These regional results strongly suggest that the FLH methodology is equally valid within similar oceanic provinces of global oceans.

Journal Article↗
PubMed2005

Effect of bio-optical parameter variability on the remote estimation of chlorophyll-a concentration in turbid productive waters: experimental results.

The analytical development and underlying hypothesis of a three-band algorithm for estimating chlorophyll-a concentration ([Chla]) in turbid productive waters are presented. The sensitivity of the algorithm to the spectral location of the bands used is analyzed. A large set of experimental observations ([Chla] varied between 4 and 217 mg m(-3) and turbidity between 2 and 78 nephelometric turbidity units) was used to calibrate and validate the algorithm. It was found that the variability of the chlorophyll-a fluorescence quantum yield and of the chlorophyll-a specific absorption coefficient can reduce considerably the accuracy of remote predictions of [Chla]. Instead of parameterizing these interferences, their effects were minimized by tuning the spectral regions used in the algorithm. This allowed us to predict [Chla] with a relative root-mean-square error of less than 30%.

Journal Article↗
PubMed2001

Cloning and characterisation of chlorophyll synthase from Avena sativa.

The chlorophyll synthase gene from oat (Avena sativa) was cloned and expressed in Escherichia coli. The deduced amino acid sequence consists of 378 amino acids including a presequence of 46 amino acids. Deletion mutants show that a core protein comprising amino acid residues 88 to 377 is enzymatically active. The sequence of the mature protein shows 85% identity with the chlorophyll synthase of Arabidopsis thaliana and 62% identity with the chlorophyll synthase of Synechocystis PCC 6803. The gene is constitutively expressed as the same transcript level is found in dark-grown and in light-grown seedlings. The enzyme requires magnesium ions for activity; manganese ions can reconstitute only part of the activity. Diacetyl and N-phenylmaleimide (NPM) inhibit the enzyme activity. Site-directed mutagenesis reveals that, out of the 4 Arg residues present in the active core protein, Arg-91 and Arg-161 are essential for the activity. Five cysteine residues are present in the core protein, of which only Cys-109 is essential for the enzyme activity. Since the wild-type and all other Cys-mutants with the exception of the mutant C304A are inhibited by N-phenylmaleimide, we conclude that the inhibitor binds to a non-essential Cys residue to abolish activity. The role of the various Arg and Cys residues is discussed.

Amino Acid Sequence↗
PubMed1999

Phosphorescence of intermediates of the terminal stage of chlorophyll biosynthesis in plants

Phosphorescence (radiative triplet state deactivation) of the intermediates involved in chlorophyll a photobiosynthesis from protochlorophyllide (Pchld) was found in greening leaves at 77K. Photoactive Pchld forms showed phosphorescence with the major spectral bands at 920 and 970 nm and 2.5-3 msec lifetime. Photochemical formation of the primary non-fluorescent intermediates (NFI) was accompanied by quenching of Pchld fluorescence and phosphorescence, phosphorescence quenching being less efficient by a factor of two. The primary chlorophyllide form Chld 684/676 that appeared as a result of NFI dark transformation showed phosphorescence with a maximum at 980 nm and 2 msec lifetime. Secondary chlorophyllide forms Chld 690/680 and Chld 695/685, the products of photochemical Chld 684/676 transformation and intermediates in biosynthesis of chlorophyll of the light harvesting complex, emitted phosphorescence with maxima at 990-995 and 1005-1010 nm and 1-2 msec lifetimes. Chlorophyll Chl 675/668, the major product of the non-photochemical "side" reaction of Chld 684/676 that probably underlies the biogenesis of photosystem II, exhibited phosphorescence with a maximum at 954-960 nm and 2 msec lifetime. Analysis of the difference spectra of the side reaction products revealed a minor 930 nm phosphorescence band that might correspond to pheophytin a formed via the "side" reaction. Triplet states of the intermediates were not quenched by carotenoids, and the quantum yields of their population at 77K were not less than 10%. Under physiological conditions the triplet intermediates might be involved in photochemical reactions, in particular, in singlet oxygen photogeneration and photodestruction of the photosynthetic apparatus.

Journal Article↗
PubMed2001

Leaf optical properties in higher plants: linking spectral characteristics to stress and chlorophyll concentration.

A number of studies have linked responses in leaf spectral reflectance, transmittance, or absorptance to physiological stress. A variety of stressors including dehydration, flooding, freezing, ozone, herbicides, competition, disease, insects, and deficiencies in ectomycorrhizal development and N fertilization have been imposed on species ranging from grasses to conifers and deciduous trees. In all cases, the maximum difference in reflectance within the 400-850 nm wavelength range between control and stressed states occurred as a reflectance increase at wavelengths near 700 nm. In studies that included transmittance and absorptance as well as reflectance, maximum differences occurred as increases and decreases, respectively, near 700 nm. This common optical response to stress could be simulated closely by varying the chlorophyll concentration of model leaves (fiberglass filter pads) and by the natural variability in leaf chlorophyll concentrations in senescent leaves of five species. The optical response to stress near 700 nm, as well as corresponding changes in reflectance that occur in the green-yellow spectrum, can be explained by the general tendency of stress to reduce leaf chlorophyll concentration.

Journal Article↗
PubMedDate not supplied

[Effect of choline chloride on photosynthetic oxygen release and variable and delayed fluorescence of chlorophyll in Chlamydomonas reinhardtii cells].

The influence of the growth retardant chlorocholine chloride on the rate of photosynthetic oxygen evolution and the induction of chlorophyll fluorescence in unicellular green algae Chlamydomonas was studied depending on concentration and the time of cell growth. It was shown that low concentrations chlorocholine chloride (0.02 g/l) affected insignificantly the photosynthesis and chlorophyll fluorescence. The growth of the culture in the presence of higher chlorocholine chloride concentrations (0.2 and 2 g/l) led to a significant reduction in the rate of oxygen production, and photoinduced changes in chlorophyll fluorescence yield. Young cells were more sensitive to chlorocholine chloride than old cells.

Animals↗
PubMed2000

A novel approach for sensitivity enhancement in atmospheric pressure chemical ionisation liquid chromatography/mass spectrometry of chlorophylls.

Differences in the ionisation efficiency of chlorophylls and their phaeophytin counterparts result in lower sensitivity for atmospheric pressure chemical ionisation mass spectrometric detection of the former. Improvement in the sensitivity of detection of chlorophyll of around an order of magnitude at a concentration of 1 x 10(-6)mol L(-1) has been achieved using post-column addition of methanoic acid during analysis by liquid chromatography/mass spectrometry (LC/MS). The method gives linear response and is a simple strategy to improve sensitivity both for LC/MS and LC/MS/MS without loss of information relating to the precise nature of the tetrapyrrole distributions. Detection levels achieved exceed those obtained by absorbance detection.

Chlorophyll↗
PubMed2005

Chlorophyll a fluorescence induction kinetics in leaves predicted from a model describing each discrete step of excitation energy and electron transfer associated with Photosystem II.

Chlorophyll a fluorescence induction (FI) is widely used as a probe for studying photosynthesis. On illumination, fluorescence emission rises from an initial level O to a maximum P through transient steps, termed J and I. FI kinetics reflect the overall performance of photosystem II (PSII). Although FI kinetics are commonly and easily measured, there is a lack of consensus as to what controls the characteristic series of transients, partially because most of the current models of FI focus on subsets of reactions of PSII, but not the whole. Here we present a model of fluorescence induction, which includes all discrete energy and electron transfer steps in and around PSII, avoiding any assumptions about what is critical to obtaining O J I P kinetics. This model successfully simulates the observed kinetics of fluorescence induction including O J I P transients. The fluorescence emission in this model was calculated directly from the amount of excited singlet-state chlorophyll in the core and peripheral antennae of PSII. Electron and energy transfer were simulated by a series of linked differential equations. A variable step numerical integration procedure (ode15s) from MATLAB provided a computationally efficient method of solving these linked equations. This in silico representation of the complete molecular system provides an experimental workbench for testing hypotheses as to the underlying mechanism controlling the O J I P kinetics and fluorescence emission at these points. Simulations based on this model showed that J corresponds to the peak concentrations of Q(-)AQB (QA and QB are the first and second quinone electron acceptor of PSII respectively) and Q(-)AQ(-)B and I to the first shoulder in the increase in concentration of Q(-)AQ(2-)B. The P peak coincides with maximum concentrations of both Q(-)AQ(2-)B and PQH2. In addition, simulations using this model suggest that different ratios of the peripheral antenna and core antenna lead to differences in fluorescence emission at O without affecting fluorescence emission at J, I and P. An increase in the concentration of QB-nonreducing PSII centers leads to higher fluorescence emission at O and correspondingly decreases the variable to maximum fluorescence ratio (F v/F m).

Chlorophyll↗
PubMed2000

Photosynthetic apparatus organization and function in the wild type and a chlorophyll b-less mutant of Chlamydomonas reinhardtii. Dependence on carbon source.

The assembly, organization and function of the photosynthetic apparatus was investigated in the wild type and a chlorophyll (Chl) b-less mutant of the unicellular green alga Chlamydomonas reinhardtii, generated via DNA insertional mutagenesis. Comparative analyses were undertaken with cells grown photoheterotrophically (acetate), photomixotrophically (acetate and HCO3-) or photoautotrophically (HCO3-). It is shown that lack of Chl b diminished the photosystem-II (PSII) functional Chl antenna size from 320 Chl (a and b) to about 95 Chl a molecules. However, the functional Chl antenna size of PSI remained fairly constant at about 290 Chl molecules, independent of the presence of Chl b. Western blot and kinetic analyses suggested the presence of inner subunits of the Chl a-b light-harvesting complex of PSII (LHCII) and the entire complement of the Chl a-b light-harvesting complex of PSI (LHCI) in the mutant. It is concluded that Chl a can replace Chl b in the inner subunits of the LHCII and in the entire complement of the LHCI. Growth of cells on acetate as the sole carbon source imposes limitations in the photon-use efficiency and capacity of photosynthesis. These are manifested as a lower quantum yield and lower light-saturated rate of photosynthesis, and as lower variable to maximal (Fv/Fmax) chlorophyll fluorescence yield ratios. This adverse effect probably originates because acetate shifts the oxidation-reduction state of the plastoquinone pool, and also because it causes a decrease in the amount and/or activity of Rubisco in the chloroplast. Such limitations are fully alleviated upon inclusion of an inorganic carbon source (e.g. bicarbonate) in the cell growth medium. Further, the work provides evidence to show that transformation of green algae can be used as a tool by which to generate mutants exhibiting a permanently truncated Chl antenna size and a higher (per Chl) photosynthetic productivity of the cells.

Animals↗
PubMed2005

Probing the lowest energy chlorophyll a states of photosystem II via selective spectroscopy: new insights on P680.

We present the wavelength dependence of homogeneous holewidths of persistent spectral holes burnt in O2-evolving Photosystem II core complexes isolated from spinach, in the temperature range 2.5-8 K. The data supports the assignment that those chlorophylls which undergo persistent spectral hole-burning are specific CP43 and CP47-trap states that transfer their excitation energy to the reaction center. The lifetime-limited holewidths show that when PS II is in the S1(QA -) (closed) state, the CP43/CP47-trap states have excited-state lifetimes in the range from 70 to 270 ps. These lifetimes correspond to excitation transfer rates to the reaction center, and are far slower than required for models in which the PS II reaction center (P680) acts as a 'shallow-trap' for excitations. For wavelengths at which both traps absorb, the hole shape is clearly a composite of two Lorentzians, corresponding to hole-burning in both states simultaneously. The temperature dependence of the homogeneous holewidth does not follow the usual T1.3 dependence found in many chlorophyll-protein systems. Our data indicates T 2 temperature dependence, typically found in crystalline systems where the chromophore is coupled to localized phonon modes.

Chlorophyll↗
PubMed1985

The relationship between light scattering and chlorophyll a fluorescence during oscillations in photosynthetic carbon assimilation.

Light scattering, which can be taken as an indicator of the transthylakoid proton gradient, and the 518-nm rise, which can be regarded as a measure of the transthylakoid membrane potential, have been followed during oscillations in chlorophyll a fluorescence, which are known to be associated with corresponding changes in photosynthetic carbon assimilation. Both components oscillated in a manner which was broadly reciprocal to chlorophyll a fluorescence. However, there was a phase shift such that the light-scattering change usually anticipated fluorescence and often also the 518-nm shift. It is concluded that the proton motive force rises and falls slightly in advance of rises and falls in carbon assimilation. The relationship of these changes to a possible underlying mechanism is discussed.

Carbon↗
PubMed1989

Enhancement of chlorophyll a fluorescence yield, low-temperature F685/F730 fluorescence emission ratio, and electron transport rate by ether phospholipids (platelet activating factor and analogs) in isolated chloroplasts.

Acetyl glyceryl ether phosphorylcholine (1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine; or platelet activating factor (PAF)), when incubated with chloroplasts or subchloroplast fractions derived from stroma or grana lamellae, induces a drastic increase in the low-temperature fluorescence emission ratio F685/F730 (77 degree K). The molecular structure requirement for the effect to be elicited is the ether bond and a long C chain at the C-1 position of glycerol, a short C chain at C-2 (or the lyso form), and a large polar head at C-3, the potent effector being PAF C-16. The effect is more pronounced in grana-derived fractions. PAF also induces an increase in the chlorophyll alpha fluorescence yield, enhances the association of chlorophyll in the supramolecular pigment-protein complexes of the thylakoid (especially those of Photosystem II), and enhances electron transfer from 1,5-diphenyl carbazide to 2,6-dichlorophenol. These effects are attributed to alteration of the Photosystem II unit organization via the incorporation/intercalation in the grana of the wedge-shaped PAF.

Chlorophyll↗
PubMed1996

Two distinct forms of the peridinin-chlorophyll a-protein from Amphidinium carterae.

Peridinin-chlorophyll a-proteins (PCPs) have been purified by combination of ammonium sulphate precipitation and cation exchange chromatography. The amino acid sequences of several of the most abundant forms have been deduced by direct protein sequencing and from DNA and indicate a highly conserved multi-gene family. At least two of the PCP genes are tandemly arranged. A novel form of the protein was also obtained in low yield with fewer peridinins (six vs eight) per chlorophyll a and with a different molecular mass (34 kDa vs 32 kDa) of its apoprotein. It had only 31% sequence identity with any of the more abundant PCP forms but retained a two-domain structure.

Amino Acid Sequence↗
PubMed1987

A new method for the separation of the constant and the variable fluorescence of chlorophyll a in in vivo.

The constant fluorescence of chlorophyll a in alga Dunaliella tertiolecta was estimated by a method of the least square regression applied to the linearized form of the equation y = axb. The value of the constant fluorescence obtained by this method was compared by the values estimated with simple linear extrapolation. Constant fluorescence, evaluated by the simple linear extrapolation of 10 ms of the initial variable fluorescence, was 50% higher than the value obtained by the method of the least square regression. We demonstrated that the estimation of constant fluorescence by the least square regression is a more correct method and provides a better comparison of results from different laboratories. This method offers a simple way to determine and separate constant fluorescence from variable fluorescence in the total yield of chlorophyll a fluorescence in "in vivo" conditions. Furthermore it facilitates the interpretation of the variable fluorescence phenomena in "in vivo".

Chlorophyll↗
PubMed1993

Derivation of the formyl-group oxygen of chlorophyll b from molecular oxygen in greening leaves of a higher plant (Zea mays).

Using mass spectroscopy, we demonstrate as much as 93% enrichment of the 7-formyl group oxygen of chlorophyll b when dark-grown, etiolated maize leaves are greened under white light in the presence of 18O2. This suggests that a mono-oxygenase is involved in the oxidation of its methyl group precursor. The concomitant enrichment of about 75% of the 13(1)-oxygen confirms the well-documented finding that this oxo group, in both chlorophyll a and b, also arises from O2. High 18O enrichment into the 7-formyl oxygen relative to the substrate 18O2 was achieved by optimization of the greening conditions in combination with a reductive extraction procedure. It indicates not only a single pathway for Chl b formyl group formation, but also unequivocally demonstrates that molecular oxygen is the sole precursor of the 7-formyl oxygen.

Chlorophyll↗
PubMed1994

Characterization of a D1-D2-cyt b-559 complex containing 4 chlorophyll a/2 pheophytin a isolated with the use of MgSO4.

A D1-D2-cyt b-559 complex containing 4 chlorophyll alpha, 1 beta-carotene and 1 cytochrome b-559 per 2 pheophytin a has been isolated from spinach with 30% yield using a Q-Sepharose Fast-Flow anion-exchange column equilibrated with 0.1% Triton X-100, 10 mM MgSO4 and 50 mM Tris-HCl (pH 7.2). The preparation was then stabilized with 0.1% dodecyl-beta-D-maltoside. This method gave a yield 10 times higher than that using a Fractogel TSK-DEAE 650(S) column equilibrated with 0.1% Triton X-100, 30 mM NaCl and 50 mM Tris-HCl (pH 7.2). The PS II RC complex was characterized using absorption and fluorescence spectroscopy at 277 and 77 K. A selective reversible bleaching under reducing conditions with maximum at 682 nm, associated with pheophytin a reduction, and light-induced absorption differences with a lifetime of 1.0 ms, ascribed to the triplet state of P680 were measured and indicated that the isolated D1-D2-cyt b-559 complex is active in charge separation. The results are compared with the data obtained for a PS II RC preparation containing 6 chlorophyll alpha, 2 beta-carotene and 1 cyt b-559 per 2 pheophytin a.

Chlorophyll↗
PubMed2004

Correlation between lifetime heterogeneity and kinetics heterogeneity during chlorophyll fluorescence induction in leaves: 1. Mono-frequency phase and modulation analysis reveals a conformational change of a PSII pigment complex during the IP thermal phase.

The relationship between the fluorescence lifetime (tau) and yield (Phi) obtained in phase and modulation fluorometry at 54 MHz during the chlorophyll fluorescence induction in dark-adapted leaves under low actinic light has been investigated. Three typical phases have been identified: (i) linear during the OI photochemical rise, (ii) convex curvature during the subsequent IP thermal rise, and (iii) linear during the PS slow decay. A similar relationship has been obtained in the fluorescence induction for the fluorescence yield measured at 685 nm plotted versus the fluorescence yield measured at 735 nm. A spectrally resolved analysis shows that the curvature of the tau-Phi relationship is not due to chlorophyll fluorescence reabsorption effects. Several other hypotheses are discussed and we conclude that the curvature of the tau-Phi relationship is due to a variable and transitory nonphotochemical quenching. We tentatively propose that this quenching results from a conformational change of a pigment-protein complex of Photosystem II core antenna during the IP phase and could explain both spectral and temporal transitory changes of the fluorescence. A variable blue shift of the 685 nm peak of the fluorescence spectrum during the IP phase has been observed, supporting this hypothesis.

Adaptation, Physiological↗