PubMed Health⌕ Search

Biomedical subjects

S Malkin

Publications and source records attributed to S Malkin.

At least 37 records · Page 2Linked to original sources
PubMed1984

Electrophotoluminescence and the electrical properties of the photosynthetic membrane. I. Initial kinetics and the charging capacitance of the membrane.

Preilluminated chloroplast membranes, and particularly hypotonically swollen vesicles (blebs), give rise to a strong characteristic luminescence (electrophotoluminescence, EPL; Ellenson and Sauer, 1976, Photochem. Photobiol., 23:113-123; Arnold and Azzi, 1971, Photochem. Photobiol., 14:233-240) during the application of a strong external electric field. A detailed kinetic study of EPL was carried out and the initial kinetics from the field onset are reported here. The fast rise time (less than 0.2 mus) of the applied external electric field together with a high instrumental time resolution allowed the observation of a characteristic delay (lag time) between the field onset and the appearance of the induced emission. The lag time decreased with increase in the applied field strength and/or the conductivity of the suspension and is interpreted to be a consequence of (a) the necessity to reach a threshold electrical potential difference in the bleb membrane, below which no emission can be triggered, and (b) the finite time required to attain such a transmembranal field during the charging process of the membrane. A quantitative analysis, connecting the lag time, the controllable experimental parameters, and the membrane electrical characteristics is presented. Its verification was carried out in both size-selected and heterogeneous bleb populations. In the latter, experiments were consistent with the assumption that the lag time reflects the charging of the largest blebs. The results indicate (a) the possibility of directly measuring the specific membrane capacitance, yielding an estimate of Cm = 1.2 +/- 0.3 microF/cm2 (the precision being particle size-homogeneity dependent); (b) A minimal transmembranal potential difference (of approximately 240 mV) is necessary to induce electrophotoluminescence; and (c) the lag duration depends on the time elapsed between the preillumination and the external field application. Correlated with the study of ionophore effects on the lag time, this suggests additivity of the light- and field-induced transmembrane potentials in attaining the threshold for emission.

Chloroplasts↗
PubMed1984

Evidence that phosphorylation and dephosphorylation regulate the distribution of excitation energy between the two photosystems of photosynthesis in vivo: Photoacoustic and fluorimetric study of an intact leaf.

State 1-state 2 transitions in an intact tobacco leaf were monitored by the photoacoustic method. Modulated oxygen evolution yield and its enhancement by continuous far-red light ("Emerson enhancement") were used to characterize the balance of light distribution between the two photosystems. These measurements were additionally supported by fluorimetry. Adaptation of the leaf to far-red light (lambda [unk] 700 nm), mainly absorbed in photosystem I (light 1), results in state 1, where short-wavelength light (light 2) is distributed in favor of photosystem II. This is shown by a low yield of oxygen evolution, a high extent of Emerson enhancement, a concomitantly high extent of fluorescence quenching by far-red light, and a low ratio of the 77 K emission peaks at 730 and 695 nm. The magnitudes of these parameters were reversed when the leaf was adapted to light 2 (state 2), indicating a change towards a more equal distribution of the excitation between the two photosystems. Preincubation of an intact leaf with NaF, a specific phosphatase inhibitor, stimulated the extent of adaptation to light 2, shown by all the above criteria, and completely abolished adaptation to light 1. Light 1 preillumination prior to NaF treatment resulted initially in state 1, but then a transition to state 2 was irreversibly induced by any light. The NaF effect was specific because NaCl did not affect the state 1-state 2 transitions. Leaching out the NaF restored the original physiological transitions of the leaf. NaF presumably acts here in the same way as it acts in isolated thylakoids-by blocking the dephosphorylation of membranal proteins (particularly the chlorophyll a/b-protein complex) phosphorylated by a light 2-activated kinase. Our results give direct support to the suggestion [Allen, J. F., Bennett, J., Steinback, K. E. & Arntzen, C. J. (1981) Nature (London) 291, 25-29] that it is the phosphorylation level of thylakoid proteins that controls the light distribution between the two photosystems in vivo, shown previously in isolated thylakoids.

Journal Article↗
PubMed1982

Functional Organization of the Chlorophyll-Containing Complexes of Chlamydomonas reinhardi: A Study of Their Formation and Interconnection with Reaction Centers in the Greening Process of the y-1 Mutant.

The stepwise synthesis and assembly of photosynthetic membrane components in the y-1 mutant of Chlamydomonas reinhardi have been previously demonstrated (Ohad 1975 In Membrane Biogenesis, Mitochondria, Chloroplasts and Bacteria, Plenum, pp 279-350). This experimental system was used here in order to investigate the process of formation and interconnection of the energy collecting chlorophylls with the reaction centers of both photosystems I and II. The following measurements were carried out: photosynthetic electron flow at various light intensities, including parts or the entire electron transfer chain; analysis of the kinetics of fluorescence emission at room temperature and fluorescence emission spectra at 77 K, and electrophoretic separation of membrane polypeptides and chlorophyll protein complexes. Based on the data obtained it is concluded that: (a) each photosystem (PSI and PSII) contains, in addition to the reaction center, an interconnecting antenna and a main or light harvesting antenna complex; (b) the formation of the light harvesting complex, interconnecting antenna, and reaction centers for each photosystem can occur independently. (c) the interconnecting antennae link the light harvesting complexes with the respective reaction centers. In their absence, energy transfer between the light harvesting chlorophylls and the reaction centers is inefficient. The formation of the interconnecting antennae and efficient assembly of photosystem components occur simultaneously with the de novo synthesis of chlorophyll and at least three polypeptides, one translated in the cytoplasm and two translated in the chloroplast. The synthesis of these polypeptides was found to be light dependent.

Journal Article↗
PubMed1981

Salt-induced microscopic changes in chlorophyll fluorescence distribution in the thylakoid membrane.

Addition of 3 mM MgCl2 to isolated pea thylakoids suspended in a medium of low osmotic strength at room temperature induces an increase in chlorophyll fluorescence similar to that observed with unswollen thylakoids. Fluorescence microscopy indicates that the MgCl2 induced increase in the emission intensity involves the formation of highly fluorescent patches on the swollen vesicles. The data seems to give additional support ton the concept that salt induced chlorophyll fluorescence changes involves the lateral movement of pigment-proteins within the thylakoid membrane in such a way as to form discrete domains.

Chlorophyll↗
PubMed1981

Photosystem II Photosynthetic Unit Sizes from Fluorescence Induction in Leaves : CORRELATION TO PHOTOSYNTHETIC CAPACITY.

The use of fluorescence induction measurements in leaves infiltrated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea has been evaluated as a routine method for estimation of the concentration of the reaction centers of photosystem II relative to total chlorophyll in a wide variety of plant species. The procedure is based on a simple theory that takes into account the attenuation of light in passing through the leaf and the linear dependence of the fluorescence induction time from different parts of the leaf on the inverse of the local light intensity. A formula to calculate the reaction center concentration of photosystem II was obtained. The effect of the light attenuation is accounted for by a correction factor which could become practically insignificant by an optimal choice of the excitation and emission wavelengths and the geometry of the photodetector with respect to the sample. Estimation of quantum yields for primary photochemistry and influence of light scattering were considered. The results demonstrate the effect of the above factors under various circumstances and are in agreement, to a first approximation, with the theory.THE UTILITY OF THE METHOD IS DEMONSTRATED BY A DETAILED STUDY OF FOUR DESERT PLANT SPECIES: estimation of reaction center concentrations of both photosystem I (by estimation of P700) and photosystem II (by the fluorescence induction method) were made and were compared to the rates of CO(2) fixation. There was a good quantitative correlation between the photosynthetic rates and the concentration of photosystem II reaction centers (expressed as per chlorophyll or per unit area of the leaf), but no such correlation was found with photosystem I reaction centers.The ratio of total chlorophyll per reaction centers II varied in the range of about 200 to 800 in different species, but there was no variation of this parameter in any single species.

Journal Article↗
PubMed1981

Photosynthetic units of sun and shade plants.

A computer analysis of fluorescence induction curves of leaves treated with 3-(3,4-dichlorophenyl)-1,1 dimethylurea was done for several species. These measurements gave the ratios of the total chlorophyll to photosystem II reaction centers. This communication is a preliminary survey of sun and shade plants and demonstrates a significant variation in this ratio. In the sun plants, the photosynthetic unit sizes (chlorophyll reaction centers) varied between 220 to 480. The shade plants gave numbers mostly in the range between 630 to 940. The computer analysis of the fluorescence data also gave the connectivity parameter of energy transfer between photosynthetic units of photosystem II which varied between 0.2 and 0.5 but did not show any obvious correlation to the photosynthetic unit size.

Journal Article↗
PubMed1980

Photoacoustic detection of photosynthetic activities in isolated broken chloroplasts.

Methodology and demonstration how to utilize the photoacoustic technique in photosynthesis research are presented. Photoacoustic signals were obtained from suspensions of isolated broken chloroplasts. In the presence of strong, continuous (non-modulated) background light the signals were normally larger than without the background light. The effect of the background light was saturable and was absent when non-active (e.g. heat-treated) samples were used, showing that the normal smaller signal in the absence of background light is a genuine reflection of the loss of heat due to the competing photochemistry. The effect of the background light is to close the reaction-centers and hence to inhibit the photochemical process. The percent difference of the photoacoustic signal (+/- background light) is taken as a measure of the photochemical activity ('photochemical loss'). Initial results demonstrate the wavelength dependence of the 'photochemical loss'. As expected there was a 'red-drop' decrease of the 'photochemical loss' for lambda greater than 690 nm, when the cofactor methyl viologen was present. Surprisingly, however, there was a 'red-rise' increase for lambda greater than 690 nm when no cofactor was present. These findings indicate that under the last conditions there is an unsuspected photoactivity of PS I which was not detected hitherto by the conventional techniques. The dependence on the background light intensity confirms this result. This photoactivity can be explained tentatively as a cyclic electron flow around PS I, present without any added cofactor. Initial results on the modulation frequency dependence in the presence of electron acceptors are also demonstrated.

Chloroplasts↗
PubMed1980

Photoacoustics in life sciences.

Photoacoustic (PA) measurements provide, by the very nature of the PA effect, the possibility to obtain information on the optical and thermal properties of samples. In addition they can yield information on the enthalpy changes and characteristic times involved in photo-induced processes as the acoustic signal is proportional to the hear produced following the absorption of the modulated excitation. In the study of optical properties the relative insensitivity to scattered light of the PA signal makes such measurement an attractive way to measure biological samples in vivo, or, at least, without the need to isolate the absorbing compounds. The dependence of the PA signal on the thermal properties of the sample is particularly useful when heterogeneous samples are studied. As a photocalorimetric method the technique shows considerable promise in the study of photo-bioenergetics, especially photosynthesis. Only in special cases can analytical applications of the PA method compete with fluorescence measurements for detection, and with increasingly sophisticated optical transmission and reflectance techniques (for identification).. However, the PA method may find important uses in fundamental research and in applied areas such as biomedicine and agricultural biochemistry.

Calorimetry↗
PubMed1979

Sulfide inhibition of photosystem II in cyanobacteria (blue-green algae) and tobacco chloroplasts.

The present study shows that in the presence of 600 nm light, sulfide acts as a specific inhibitor of photosynthetic electron transport between water and Photosystem II in the cyanobacteria Aphanothece halophytica and Synechococcus 6311 as well as in tobacco chloroplasts. In the presence of 600 nm light sulfied affects the fast fluorescence transients as does a low concentration (10 mM) of hydroxylamine; the fluorescence yield decreases in the presence of either chemical and can be restored by the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. In chloroplasts, however, NH2OH, an electron donor at high concentrations (40 mM), relieves the sulfide effect. In the dark, sulfide affects the cyanobacterial fluorescence transients through decrease of oxygen tension. The fluorescence yield increases in a similar pattern to that observed under nitrogen flushing. Upon omission of sulfide in A. halophytica, the characteristic aerobic fluorescence transients return, consistent with the ease of alternation between oxygenic and sulfide-dependent anoxygenic photosynthesis in many cyanobacteria.

Aerobiosis↗
PubMed1979

Anomalous oxygen uptake from isolated chloroplasts inhibited in photosystem II and without external electron donors.

Light induced modulated signal of oxygen uptake by isolated chloroplasts in the presence of methyl viologen, when Photosystem II activity was inhibited and in the absence of any electron donors, was detected by a modulated oxygen Pt electrode, polarized negatively. Evidence is brought to show that an electrochemical process which takes place on the surface of the negatively polarized Pt-cathode produces an intermediate which serves as an electron donor to Photosystem I. Atempts to identify this intermediate show that it may be very probably the superoxide radical generated by the electrochemical reduction of oxygen which continuously diffuses from the external circulating medium to the electrode.

Cell Membrane↗
PubMed1979

Cold storage of isolated class C chloroplasts: optimal conditions for stabilization of photosynthetic activities.

Preservation of photosynthetic activities (photophosphorylation, electron transport, fluorescence induction, 0.3-second delayed light emission) of isolated broken (class C) chloroplasts by low temperature storage was investigated under a wide range of conditions in order to optimize long time activity retention.The more labile functions (photophosphorylation and electron transport) required very low temperatures (below -79 C) and relatively high (above 20%, v/v) concentrations of cryoprotectives for satisfactory stabilization. Fluorescence induction and delayed light emission were less sensitive, especially during the 1st month of storage.Taking into account the effect of cryoprotectives on absolute activities prior to freezing, optimum activity retention was observed with a medium containing ethylene glycol (30%, v/v) and a storage temperature of -100 C or below. In this case, given fast thawing and high chloroplast concentration, practically 100% preservation of all of the photosynthetic activities investigated was obtained for at least 10 months, even with very simple freezing and storage procedures.The same optimal medium at somewhat higher temperatures (-79 C and to a lesser extent at -41 C) caused a dramatic uncoupling effect: photophosphorylation was inhibited in a few hours, while electron transport increased 3- to 5-fold. The enhanced electron transport was stable for almost a month and then declined sharply. This uncoupling effect was specific only to ethylene glycol.

Journal Article↗
PubMed1978

Induction patterns of delayed luminescence fromisolated chloroplasts. I. Response of delayed luminescence to changes in the prompt fluorescence yield.

1. Using a phosphoroscope, delayed luminescence and prompt chlorophyll fluorescence from isolated chloroplasts have been compared during the induction period. 2. Two distinct decay components of delayed luminescence were measured a "fast" component (from approximately 1 ms to approximately 6 ms) and a "slow" component (at approximately 6 ms). 3. The fast luminescence component often did not correlate with the fluorescence changes while the slow component significantly changed its intensity during the induction period in a manner which could usually be linearly correlated with variable portion of the fluorescence yield change. 4. This correlation was evident after preillumination with far-red light or after allowing a considerable time for dark relaxation. 5. The close relationship between the slow luminescence component and variable fluorescence yield was observed with a large range of light intensities and also in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea which considerably changes the fluorescence induction kinetics. 6. Valinomycin and other antibiotics reduced the amplitude of the 6 ms (slow) luminescence without affecting its relation with the fluorescence induction suggesting possibly that a constant electrical gradient exist in the dark or formed very rapidly in the light, which effects the emission intensity. 7. Changes in salt levels of suspending media equally affected the amplitude of both delayed luminescence and variable fluorescence under conditions when the reduction of Q is maximal and constant. 8. The results are discussed in terms of several models. It is concluded that the model of independent Photosystem II units together with photosynthetic back reaction concept is incompatible with the data. Other alternative models (the "lake" model and photosynthetic back reaction; recombination of charges in the antenna chlorophyll; the "w" hypothesis) were in closer agreement with the results.

Chloroplasts↗
PubMed1978

Development and Repair of Photosystem II Activity in Normal and Chloramphenicol-treated Euglena gracilis Cells.

Photosystem II activity, development, and organization were studied in membranes from dark-adapted Euglena gracilis Klebs var. Z Pringsheim cells during a modulated greening process of light-dark-light cycles. The results obtained from measurements of overlapping partial photosystem II (PSII) reactions (fluorescence induction parameters, quantum yield, flash yield, and maximal rate of H(2)O --> 2,6-dichlorophenolindophenol [DCIP] and 1,5-diphenylcarbazide [DPC] --> DCIP reactions) during these cycles indicate the formation of active PSII units in the dark. The necessity for proteins from the chloroplast translational machinery for this formation is evidenced by the inhibition of synthesis of the PSII units in chloramphenicol-treated cells. The effect of this drug, both during the dark and second light periods, can be summarized as follows: (a) disruption of the electron transfer connection to the plastoquinone pool, or decrease in the pool size; (b) loss of excitation energy transfer efficiency in the second light period; (c) impairment of the O(2) evolution appratus, as shown by comparison of the efficiency of DCP and H(2)O as electron donors.These conclusions are based on the use of a previously developed method of measurement and analysis of data (Cahen et al. 1976 Plant Physiol 58: 257-267).

Journal Article↗