RESUMEN
The concept that the Carnot efficiency places an upper limit of 0.60-0.75 on the thermodynamic efficiency of photosynthetic primary photochemistry is examined using the PSI-LHCI preparation. The maximal quantum efficiency was determined approximately 0.99 which yielded a thermodynamic efficiency of 0.96, a value far above that predicted on the basis of the Carnot efficiency. The commonly presented reasoning leading to the Carnot efficiency idea was therefore critically examined. It is concluded that the crucial assumption that the pigment system, under illumination, is in equilibrium with the incident light field, at a black body temperature of Tr, is erroneous, as the temperature of the excited state pigments was experimentally shown to be that of the sample solvent (thermal bath), 280 K in this case. It is concluded that the classical reasoning used to describe the thermodynamics of heat systems is not applicable to "photonic" systems such as plant photosystems.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/química , TermodinámicaRESUMEN
In spinach thylakoids, the quenching of the singlet excited state in the photosystem II antenna by m-dinitrobenzene does not change the rate of the light induced degradation of the D1 reaction centre protein and offers only limited protection against photoinhibition itself. These results are discussed in terms of the role of non-photochemical quenching as a photoprotective strategy.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Tilacoides/química , Dinitrobencenos/química , Fluorescencia , Immunoblotting , Luz , Fotoquímica , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Proteínas del Complejo del Centro de Reacción Fotosintética/inmunología , Complejo de Proteína del Fotosistema II , Spinacia oleracea/química , Tilacoides/metabolismoRESUMEN
Selective destruction of the strongly dichroic red-shifted chlorophyll form (C709 nm) in photosystem I (PSI) trimers from Spirulina, by either non-selective high intensity illumination (photobleaching) or incubation with low concentrations of Triton X-100 is accompanied by changes in the circular dichroism spectrum of the same amplitude and of opposite sign at 677 nm. The data are interpreted in terms of a dimeric chlorophyll structure with excitonic bands at these two wavelengths. Similar photobleaching experiments with PSI-200 from maize also suggest the presence of bulk antenna/red form excitonic interactions.
Asunto(s)
Clorofila/metabolismo , Dicroismo Circular , Cianobacterias/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Clorofila/química , Color , Dimerización , Luz , Complejos de Proteína Captadores de Luz , Octoxinol/farmacología , Complejo de Proteína del Fotosistema I , Unión Proteica/efectos de los fármacos , Unión Proteica/efectos de la radiación , Zea mays/químicaRESUMEN
Evidence is presented, by means of both fluorescence and action spectroscopy, that a small, spectroscopically heterogeneous population of both Chl a and Chl b molecules is present in isolated spinach thylakoids and is active in photoinhibition. The broadness of the action spectrum suggests that degraded or incompletely assembled pigment-protein complexes may be involved.
Asunto(s)
Clorofila/metabolismo , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Hojas de la Planta/fisiología , Spinacia oleracea/fisiología , Tilacoides/fisiología , Clorofila A , Luz , Complejos de Proteína Captadores de Luz , Espectrometría de Masas , Hojas de la Planta/metabolismo , Espectrometría de Fluorescencia , Spinacia oleracea/metabolismo , Tilacoides/metabolismoRESUMEN
In photosystem I trimers of Spirulina platensis a major long wavelength transition is irreversibly bleached by illumination with high-intensity white light. The photobleaching hole, identified by both absorption and circular dichroism spectroscopies, is interpreted as the inhomogeneously broadened Q(y) transition of a chlorophyll form that absorbs maximally near 709 nm at room temperature. Analysis of the mean square deviation of the photobleaching hole between 80 and 300 K, in the linear electron-phonon frame, indicates that the optical reorganization energy is 52 cm(-1), four times greater than that for the bulk, short-wavelength-absorbing chlorophylls, and the inhomogenous site distribution bandwidth is close to 150 cm(-1). The room temperature bandwidth, close to 18.5 nm, is dominated by thermal (homogeneous) broadening. Photobleaching induces correlated circular dichroism changes, of opposite sign, at 709 and 670 nm, which suggests that the long wavelength transition may be a low energy excitonic band, in agreement with its high reorganization energy. Clear identification of the 709-nm spectral form was used in developing a Gaussian description of the long wavelength absorption tail by analyzing the changing band shape during photobleaching using a global decomposition procedure. Additional absorption states near 720, 733, and 743 nm were thus identified. The lowest energy state at 743 nm is present in substoichiometric levels at room temperature and its presence was confirmed by fluorescence spectroscopy. This state displays an unusual increase in intensity upon lowering the temperature, which is successfully described by assuming the presence of low-lying, thermally populated states.
Asunto(s)
Cianobacterias/fisiología , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Clorofila/química , Dicroismo Circular , Luz , Fotoquímica , Proteínas del Complejo del Centro de Reacción Fotosintética/efectos de la radiación , Conformación Proteica , Espectrofotometría , TermodinámicaRESUMEN
A photosystem I preparation from maize, containing its full antenna complement (PSI-200) and in which detergent effects on chlorophyll coupling are almost completely absent, has been studied by time-resolved fluorescence techniques with approximately 5 ps resolution at 280 and 170 K in the wavelength interval of 690-780 nm. The data have been analyzed in terms of both the decay-associated spectra (DAS) and the time-resolved emission spectra (TRES). As in a previous room temperature study [Turconi, S., Weber, N., Schweitzer, D., Strotmann, H., and Holzwarth, A. R. (1994) Biochim. Biophys. Acta 1187, 324-334], the 280 K decay is well described by three DAS components in the 11-130 ps time range, the fastest of which displays both positive and negative amplitudes characteristic of excitation transfer from the bulk to the red antenna forms. Both the 57 and 130 ps components have all positive amplitudes and describe complex decay and equilibration processes involving the red forms. At 170 K, four major components in the 10-715 ps time range are required to describe the decay. The fastest represents bulk to red form transfer processes, while the 55, 216, and 715 ps decays, with all positive amplitudes, have maxima near 720, 730, and 740 nm, respectively, in accord with previous steady-state fluorescence measurements. The width and asymmetry of these DAS indicate that they are spectrally complex and represent decay and equilibration processes involving the red forms. Spectral evolution during the fluorescence decay process was analyzed in terms of the TRES. The red shifting of the TRES was analyzed in terms of the first central spectral moment (mean spectral energy) which is biexponential at both temperatures. The slower component, which describes equilibration between the red forms, leads to spectral red shifting during the entire fluorescence decay process, and the mean lifetimes of the spectral moments at 280 and 170 K (86 and 291 ps, respectively) are similar to the mean lifetimes of the fluorescence decays (119 and 384 ps, respectively). Thus, both spectral evolution and the trapping-associated fluorescence decay occur on a similar time scale, and both processes display a very similar temperature sensitivity. On the basis of these data, it is concluded that trapping in PSI-200 is to a large extent rate-limited by excitation diffusion in the antenna and in particular by the slow "uphill" transfer from the low-energy forms to the bulk and/or inner core chlorophyll molecules.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/química , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Zea mays/metabolismo , Cinética , Complejos de Proteína Captadores de Luz , Modelos Teóricos , Complejo de Proteína del Fotosistema I , Espectrometría de Fluorescencia , Espectrofotometría , Factores de TiempoRESUMEN
The spectral characteristics of fluorescence quenching by open reaction centres in isolated Photosystem II membranes were determined with very high resolution and analysed. Quenching due to photochemistry is maximal near 687 nm, minimal in the chlorophyll b emission interval and displays a distinctive structure around 670 nm. The amplitude of this 'quenching hole' is about 0.03 for normalised spectra. On the basis of the absorption spectra of isolated chlorophyll-protein complexes, it is shown that these quenching structures can be exactly described by assuming that photochemistry lowers the fluorescence yield of the reaction centre complex (D1/D2/cytb (559)) plus CP47, with quenching of the former complex being approximately double that of the latter complex. These data, which qualitatively indicate that there are kinetically limiting processes for primary photochemistry in the antenna, have been analysed by means of several different kinetic models. These models are derived from present structural knowledge of the arrangement of the chlorophyll-protein complexes in Photosystem II and incorporate the reversible charge separation characteristic of the exciton/radical pair equilibration model. In this way it is shown that Photosystem II cannot be considered to be purely trap limited and that exciton migration in the antenna imposes a diffusion limitation of about 30%, irrespective of the structural model assumed.
RESUMEN
The red tail of the absorption spectrum of the D1-D2-cytb559 complex, defined as the absorption signal not described by the two Gaussian sub-bands associated with the intense electronic transitions at 680 and 683 nm, exhibits anomalous temperature behavior. This tail was analyzed in the temperature interval between 80 and 300 K in terms of the mean square deviation (sigma2) of the total Qy absorption band and by Gaussian sub-band decomposition. The value of the average optical reorganization energy (Snum) obtained from the temperature dependence of sigma2 for the whole absorption band was 32 cm(-1), and changed to 16-20 cm(-1) after subtraction of the sub-bands describing the red tail. This latter value is in agreement with the hole burning literature data for chlorophyll bound to proteins, and indicates that the rather high value for the apparent optical reorganization energy obtained by analysis of the total Qy band of the D1-D2-cytb559 complex is determined by the temperature sensitivity of the red tail. This suggests that the long wavelength absorption tail might be due to vibrational transitions associated with vibrational modes in the range of 80-150 cm(-1) which are thermally accessible and give rise to an absorption signal on the low-energy side of the (0,0) transition. On the basis of this assumption, the electron-phonon coupling strength (S) for these modes is estimated to be in the range 0.028-0.18. This interpretation furthermore supports the idea that the electronic transition near 683 nm is that of a monomer chlorophyll.
Asunto(s)
Fragmentos de Péptidos/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Absorción , Grupo Citocromo b/química , Complejos de Proteína Captadores de Luz , Modelos Químicos , Distribución Normal , Feofitinas/química , Complejo de Proteína del Fotosistema II , Espectrofotometría , TemperaturaRESUMEN
One of the strains of the marine green alga Ostreobium sp. possesses an exceptionally large number of long wavelength absorbing chlorophylls (P. Haldall, Biol. Bull. 134, 1968, 411-424) as evident from a distinct shoulder in the absorption spectrum at around 710 nm while in the other strain this shoulder is absent. Therefore, Ostreobium offers a unique possibility to explore the origin of these red-shifted chlorophylls, because strains with and without these spectral forms can be compared. Here, we characterize these red forms spectroscopically by absorption, fluorescence and CD spectroscopy. In the CD spectra at least three spectroscopic red forms are identified which lead to an unusual room temperature fluorescence spectrum that peaks at 715 nm. The gel electrophoretic pattern from thylakoids of Ostreobium sp. shows an intense band at 22 kDa which correlates with the presence or absence of long wavelength absorbing pigments. By protein sequencing of the N-terminus of the 22-kDa polypeptide and sequence alignments, this was identified as an Lhca1-type light-harvesting complex. The abundance of this polypeptide - and a possibly co-migrating one - in Ostreobium sp. indicates an antenna size of approximately 340 chlorophyll molecules (Chl a and Chl b) per PS IIalpha reaction center, which is significantly larger than in higher plants ( approximately 240). The red forms are more abundant in the interior of the thalli where a 'shade-light' light field is expected than in the white-light exposed surface. This demonstrates that algae exist which may be able to up-regulate the synthesis of large amounts of LHCI and associated red forms under appropriate illumination conditions.
Asunto(s)
Proteínas de Arabidopsis , Clorofila/química , Chlorophyta/química , Complejos de Proteína Captadores de Luz , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Complejo de Proteína del Fotosistema I , Proteínas de Plantas/química , Secuencia de Aminoácidos , Chile , Proteínas de Unión a Clorofila , Chlorophyta/genética , Dicroismo Circular , Electroforesis en Gel de Poliacrilamida , Datos de Secuencia Molecular , Mar del Norte , Alineación de Secuencia , Espectrometría de Fluorescencia , EspectrofotometríaRESUMEN
The intact photosystem I of maize containing its full antenna complement (PSI-200) has been purified and fractionated into the core and outer antenna (LHCI) components. It is demonstrated by absorption and fluorescence spectroscopy that at least 80% of the long wavelength absorbing antenna pigments (red forms) are located in LHCI. Absorption spectra in the Qy region of all three preparations were measured between 72 and 300 K and subjected to a thermal broadening analysis. Data are interpreted in the linear electron-phonon coupling assumption, and the average optical reorganization energy (Snum) for the bulk pigment band and the red absorption tail determined. A marked asymmetry in Snum values across the absorption band is demonstrated. The bulk pigments in all three preparations have rather low values, in the range of 15-25 cm-1, suggesting that Stokes shifts for the absorption forms are in the 1. 5-3 nm range. On the other hand the red forms have markedly greater reorganization energies. While a direct thermal analysis of the red tail indicates minimum Snum values of around 60 cm-1, when the contribution of the red tail of the bulk pigments is corrected for in LHCI, the more reliable value of 110 cm-1 is obtained. These high Snum values for the red pigment forms suggest that they have unusually wide homogeneously broadened absorption bands and large Stokes shifts (6-11 nm).
Asunto(s)
Clorofila/química , Complejos de Proteína Captadores de Luz , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Complejo de Proteína del Fotosistema II , Proteínas de Plantas , Proteínas Portadoras/química , Cloroplastos/química , Complejo de Proteína del Fotosistema I , Espectrometría de Fluorescencia , Espectrofotometría , Temperatura , Termodinámica , Zea maysRESUMEN
The minor photosystem II antenna complex CP29(Lhcb-4) has been reconstituted in vitro with the Lhcb-4 apoprotein, overexpressed in Escherichia coli, and the native pigments. Modulation of the pigment composition during reconstitution yields binding products with markedly different chlorophyll a/b binding ratios even though the total number of bound chlorophylls (a plus b) remains constant at eight. A thermodynamic analysis of steady state absorption and fluorescence spectra demonstrates that all chlorophylls are energetically coupled, while the kinetics of chlorophyll photooxidation indicate that triplet chlorophyll-carotenoid coupling is also conserved during pigment binding in vitro. The influence of the chlorophyll a/b binding ratio on the absorption spectra measured at 72 and 300 K is analyzed for the Qy absorption region. Increased chlorophyll b binding leads to large increases in absorption in the 640-660 nm region, while absorption in the 675-690 nm interval decreases markedly. These changes are analyzed in terms of a Gaussian decomposition description in which the eight subbands display a temperature-dependent broadening in agreement with the weak electron-phonon coupling demonstrated for other antenna chlorophyll spectral forms. In this way, we demonstrate that increased chlorophyll b binding leads to increased absorption intensity associated with the subbands at 640, 648, 655, and 660 nm and decreased intensity for the long wavelength subbands at 678 and 684 nm. The wavelength position of all subbands is unchanged. The above data are interpreted to indicate that CP29 has eight chlorophyll binding sites, many or all of which can be occupied by either chlorophyll a or chlorophyll b according to the conditions in which pigment binding occurs. Chlorophyll b absorption is primarily associated with four subbands located at 640, 648, 655, and 660 nm. The invariance of the wavelength position of the absorption bands in recombinant products with different chlorophyll a/b binding stoichiometries is discussed in terms of the mechanism involved in the formation of spectral bands. We conclude that pigment-protein interactions dominate in the determination of spectral heterogeneity with probably only minor effects on absorption associated with pigment-pigment interactions.
Asunto(s)
Clorofila/metabolismo , Complejos de Proteína Captadores de Luz , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Complejo de Proteína del Fotosistema II , Apoproteínas/química , Apoproteínas/metabolismo , Sitios de Unión , Clorofila A , Clonación Molecular , Escherichia coli , Cinética , Luz , Oxidación-Reducción , Pigmentos Biológicos/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Espectrometría de Fluorescencia , EspectrofotometríaRESUMEN
Absorption spectra in the Qy region of the light harvesting complex II (LHCII) have been measured in the temperature range 70-300 K. The spectra were analyzed by evaluating the temperature dependence (a) of the total bandwidth and (b) of the sub-bands obtained by numerical decomposition in terms of double Gaussians. The thermal broadening of the bands are interpreted, in both cases, as a homogeneous component, due to the presence of linear electron-phonon coupling, plus an inhomogeneous component, due to both statistical energy fluctuations at each pigment site and heterogeneity of the sample itself. Sub-bands analysis, in which eight major components are identified, yields a reorganization energy 9 cm(-1) < or = Svm < or = 14 cm(-1) and an inhomogeneous contribution in the range 120-170 cm(-1). In all cases the bands are substantially symmetrical in the 70-300 K temperature range. This observation gains theoretical support from an analysis of the band moments, when the influence of a low-frequency vibrational mode is considered. Analysis of the total absorption band yields Svm approximately 70 cm(-1); however, this high value is reduced to Svm approximately 11-20 cm(-1) when the red-most sub-band, with maximum at 684 nm, is eliminated at all temperatures. These data are discussed in terms of the underlying transitions, giving strong support to the presence of extreme red absorption bands in LHCII. The presence of another low-frequency mode with vm > 20-30 cm(-1) is also proposed.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/química , Cinética , Modelos Químicos , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Espectrofotometría , Temperatura , TermodinámicaRESUMEN
Photosystem I with its full antenna complement (PSI-LHCI) has been prepared by mild detergent solubilization with octyl beta-D-glucopyranoside from maize thylakoids. A preliminary polypeptide analysis is presented. At room temperature, the steady-state fluorescence derives from an almost perfectly thermalized state, as demonstrated by a Stepanov analysis, in which about 90% of the excited states are associated with the red chlorophyll spectral forms absorbing above 700 nm. Equilibration is temperature-sensitive and is lost at T < 200 K. A careful analysis of fluorescence between 75 and 280 K clearly demonstrates the presence of at least three red chlorophyll spectral forms with emission maxima at 720, 730, and 742 nm, the absorption origin bands of which have been calculated at 714, 725, and 738 nm. On the basis of a minor deviation from thermal equilibration around 695 nm, it is suggested that at least 3-4 antenna chlorophylls, with an average absorption near 695 nm, are strongly coupled to P700. Thermodynamic analysis of absorption and fluorescence spectra indicates that the equilibrium, absorption-weighted excited state population of the P700 dimer is around 0.013 assuming that the low-energy exciton state possesses all the oscillator strength. The average free energy for excitation transfer from antenna to P700 is thus calculated to be -0.26 kT at room temperature. This indicates that P700 is almost isoenergetic with its antenna at room temperature when the red forms are taken fully into account. From the calculated excited state population of P700, we estimate that the primary charge separation rate in PSI is 1-2 ps-1.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/química , Zea mays/química , Complejos de Proteína Captadores de Luz , Complejo de Proteína del Fotosistema I , Espectrometría de Fluorescencia , TermodinámicaRESUMEN
Photosystem II, which has a primary photochemical charge separation time of about 300 ps, is the slowest trapping of all photosystems. On the basis of an analysis of data from the literature this is shown to be due to a number of partly independent factors: a shallow energy funnel in the antenna, an energetically shallow trap, exciton dynamics which are partly 'trap limited' and a large antenna. It is argued that the first three of these properties of Photosystem II can be understood in terms of protective mechanisms against photoinhibition. These protective mechanisms, based on the generation of non photochemical quenching states mostly in the peripheral antenna, are able to decrease pheophytin reduction under conditions in which the primary quinone, QA, is already reduced, due to the slow trapping properties. The shallow antenna funnel is important in allowing quenching state-protective mechanisms in the peripheral antenna.
RESUMEN
Absorption spectra of the isolated D1/D2/cytochrome b-559 complex have been measured in the temperature range 80-300 K. All spectra were analyzed in terms of a linear combination of Gaussian bands and the thermal broadening data interpreted in terms of a model in which the spectrum of each pigment site is broadened by (a) a homogeneous component due to linear electron-phonon coupling to a low-frequency protein vibration and (b) an inhomogeneous component associated with stochastic fluctuations at each pigment site. In order to obtain a numerically adequate description of the absorption spectra, a minimum number of five sub-bands is required. Further refinement of this sub-band description was achieved by taking into account published data from hole burning and absorption difference spectroscopy. In this way, both a six sub-band description and a seven sub-band description were generated. In arriving at the seven sub-band description, the original five sub-band wavelength positions were essentially unchanged. Thermal broadening analysis of the seven sub-band description yielded data which displayed the closest correspondence with the literature observations. The wavelength positions of the sub-bands were near 661, 667, 670, and 675 nm, with two bands near 680 and 684 nm. The two almost isoenergetic sub-bands near 680 nm, identified as P680 and pheophytin, have optical reorganization energies around 40 and 16 cm-1, respectively. All other sub-bands, identified as accessory pigments, have optical reorganization energies close to 16 cm-1.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Grupo Citocromo b/química , Calor , Complejo de Proteína del Fotosistema II , Espectrofotometría , Fenómenos Químicos , Química Física , Cloroplastos/química , Complejos de Proteína Captadores de Luz , Matemática , Feofitinas/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Zea maysRESUMEN
Room temperature and 10 K absorption and linear dichroism spectra of the chlorophyll-protein complexes comprising the outer antenna of PSII (LHCII, CP29, CP26, CP24) have been analyzed in terms of a linear combination of asymmetric Gaussian bands. The results demonstrate the following: (a) The absorption and linear dichroism spectra of each sample can be described by nearly the same set of Gaussian bands at room temperature and 10 K. (b) The relative distributions of the transition moments of the major red-absorbing spectral forms seem to be similar in all four outer antenna chlorophyll-protein complexes at room temperature, with the 684-nm band being oriented closest to the particle plane at room temperature and the 677- and 669-nm bands being tilted at progressively greater angles out of the particle plane. The shorter wavelength transitions seem to be oriented close to the magic angle, but interpretation is complicated in this spectral region due to the low linear dichroism values and by overlap with vibrational bands. (c) The 684-nm band, detected in room temperature absorption and linear dichroism spectra of all complexes, vanishes at 10 K.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/química , Clorofila/química , Complejos de Proteína Captadores de Luz , Complejo de Proteína del Fotosistema II , Proteínas de Plantas/química , Análisis Espectral , Zea maysRESUMEN
A study of the absorption and fluorescence characteristics of the D1/D2/cytb-559 reaction centre complex of Photosystem II has been carried out by gaussian decomposition of absorption spectra both at room temperature and 72 K and of the room temperature fluorescence spectrum. A five component fit was found in which the absorption and fluorescence sub-bands could be connected by the Stepanov relation. The photobleaching and light-activated degradation in the dark of long wavelength pigments permitted a further characterisation of the absorption bands. The absorption (fluorescence) maxima of the five bands at room temperature are 660 nm (670 nm), 669 nm (675 nm), 675 nm (681 nm), 680 nm (683 nm), 681 nm (689 nm). A novel feature of this analysis is the presence of two approximately isoenergetic absorption bands near 680 nm at room temperature. The narrower one (FWHM=12.5 nm) is attributed to pheophytin while the broader band (FWHM=23 nm) is thought to be P680. The P680 band width is discussed in terms of homogeneous and site inhomogeous band broadening. The P680 fluorescence has a large Stokes shift (≈9 nm) and most fluorescence in the 690-700 nm range is associated with this chromophore.The three accessory pigment bands are broad (FWHM=17-24 nm) and the 660 nm gaussian is largely temperature insensitive thus indicating significant site inhomogeneous broadening.The very slight narrowing of the D1/D2/cytb-559 Qy absorption at crytogenic temperatures is discussed in terms of the coarse spectral inhomogeneity associated with the spectral forms and the apparently large site inhomogeneous broadening of short wavelength accessory pigments.
RESUMEN
The chlorophyll-protein complexes that form the antenna system of photosystem II have been purified and analyzed in terms of the commonly observed chlorophyll spectral forms. With the exception of chlorophyll b, which is known to be associated with the complexes comprising the outer antenna (LHCII, CP24, CP26, CP29), the spectral forms occur with similar absorption maxima and are present in rather similar amounts in each of the antenna complexes. On the basis of the published chlorophyll stoichiometries for the complexes in photosystem II antenna, the distribution of the spectral forms in a "reconstituted" antenna has been determined. These data were used to calculate the equilibrium population of excited states within the various chlorophyll-protein complexes within photosystem II. This was compared with the light absorption capacity of each of the complexes in the "reconstituted" antenna. The ratio of these two parameters (excited-state equilibrium distribution/absorption capacity) was determined to be 1.21 for the inner (core) antenna and 0.88 for LHCII. The standard free energy change for exciton transfer from the outer to the inner antenna was calculated to be -0.17 kcal mol-1. It is concluded that the photosystem II antenna is arranged as a very shallow funnel.
Asunto(s)
Clorofila/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Transferencia de Energía , Glucósidos/farmacología , Complejos de Proteína Captadores de Luz , Complejo de Proteína del Fotosistema II , Análisis Espectral , Zea maysRESUMEN
Irradiation of the principal photosystem II light-harvesting chlorophyll-protein antenna complex, LHC II, with high light intensities brings about a pronounced quenching of the chlorophyll fluorescence. Illumination of isolated thylakoids with high light intensities generates the formation of quenching centres within LHC II in vivo, as demonstrated by fluorescence excitation spectroscopy. In the isolated complex it is demonstrated that the light-induced fluorescence quenching: a) shows a partial, biphasic reversibility in the dark; b) is approximately proportional to the light intensity; c) is almost independent of temperature in the range 0-30°C; d) is substantially insensitive to protein modifying reagents and treatments; e) occurs in the absence of oxygen. A possible physiological importance of the phenomenon is discussed in terms of a mechanism capable of dissipating excess excitation energy within the photosystem II antenna.
RESUMEN
An experimental analysis is presented concerning the effect on relative light absorption by the two photosystems caused by (a) a highly light scattering environment (the "detour effect") and (b) light filtration across successive chloroplast layers (the "light attenuation effect"). Both suspensions of isolated chloroplasts and leaves were employed.It is concluded that within a single spinach leaf these phenomena are likely to lead to only rather small increases in relative photosystem I absorption and activity with respect to photosystem II and will thus not exert a significant effect on non cyclic electron transport. On the contrary when light is filtrated across successive vegetation layers (shade light) significant increases in the relative PSI absorption and activity may be encountered.It is determined that the "detour effect" in mature leaves from a variety of plants increases overall photosynthetically useful light absorption by 35-40%.