Photosynthetic free energy transduction related to the electric potential changes across the thylakoid membrane

A model based on our present knowledge of photosynthetic energy transduction is presented. Calculated electric potential profiles are compared with microelectrode recordings of the thylakoid electric potential during and after actinic illumination periods of intermediate duration. The information content of the measured electric response is disclosed by a comparison of experimental results with calculations. The proton flux through the ATP synthase complex is seen to markedly influence the electric response. Also the imbalance in maximum turnover rate between the two photosystems, common to obligate shade plants like Peperomia metallica used in the microelectrode experiments, is clearly reflected in the electric potential profile.Dedicated to Prof. L.N.M. Duysens on the occasion of his retirement.     

Identification and modulation of a voltage-dependent anion channel in the plasma membrane of guard cells by high-affinity ligands.

Guard cell anion channels (GCAC1) catalyze the release of anions across the plasma membrane during regulated volume decrease and also seem to be involved in the targeting of the plant growth hormones auxins. We have analyzed the modulation and inhibition of these voltage-dependent anion channels by different anion channel blockers. Ethacrynic acid, a structural correlate of an auxin, caused a shift in activation potential and simultaneously a transient increase in the peak current amplitude, whereas other blockers shifted and blocked the voltage-dependent activity of the channel. Comparison of dose-response curves for shift and block imposed by the inhibitor, indicate two different sites within the channel which interact with the ligand. The capability to inhibit GCAC1 increases in a dose-dependent manner in the sequence: probenecid less than A-9-C less than ethacrynic acid less than niflumic acid less than IAA-94 less than NPPB. All inhibitors reversibly blocked the anion channel from the extracellular side. Channel block on the level of single anion channels is characterized by a reduction of long open transitions into flickering bursts, indicating an interaction with the open mouth of the channel. IAA-23, a structural analog of IAA-94, was used to enrich ligand-binding polypeptides from the plasma membrane of guard cells by IAA-23 affinity chromatography. From this protein fraction a 60 kDa polypeptide crossreacted specifically with polyclonal antibodies raised against anion channels isolated from kidney membranes. In contrast to guard cells, mesophyll plasma membranes were deficient in voltage-dependent anion channels and lacked crossreactivity with the antibody.     

Should we expect strange attractors behind plankton dynamics - and if so, should we bother?

Chaotic behaviour can easily be generated from simple deterministicmodels of a number of ecological interactions. It is arguedthat these interactions, as well as the conditions necessaryto produce chaos from them, are common in planktonic communitiesFurthermore, attention is drawn to the fact that sustained algalfluctuations are observed in micro-ecosystems that are keptunder constant external conditions for as long as 10 years.These fluctuations are shown to have the same characteristicsas those arising from a simple chaotically behaving model ofcompeting algae. Recently, signs of chaos have also been shownin a series of plankton data from the field. It is concludedthat there are, in fact, several independent reasons to believethat planktonic systems generally have a strange attractor,rather than a static or cyclic equilibrium. The chaotic naturehas some implications that may warrant a reconsideration ofour framework for thinking about plankton dynamics. For instance'(i) the observed fluctuations can be seen as a special typeof equilibrium, the nature of which can be understood from therelationships within the system; and (ii) although the rangeof possible behaviour of the system is well defined, the actualcourse of events is unpredictable, because small differencesin initial condition expand in time exponentially. The conclusionsfrom chaos theory sound quite spectacular, however, many ofthem do not appear to be essentially new to plankton ecologyIt is argued that the discrepancy between the mathematical findingsand present-day knowledge of the mechanisms behind planktondynamics are to a large extent semantic. Nonetheless, the seeminglysmall step of accepting that these systems are fundamentallychaotic implies the acceptance of the fact that we will neverbe able to predict their long-term behaviour accurately     

Calcium-calmodulin signalling is involved in light-induced acidification by epidermal leaf cells of pea, Pisum sativum L.

Pathways of signal transduction of red and blue light-dependentacidification by leaf epidermal cells were studied using epidermalstrips of the Argenteum mutant of Pisum sativum. In these preparationsthe contribution of guard cells to the acidification is minimal.The hydroxypyridine nifedipine, a Ca²⁺-channel blocker, partlyinhibited the response to both blue and red light, while thephenylalkylamine, verapamil, a Ca²⁺-channel blocker that hasbeen shown in plant cells also to block K⁺-channels, causednearly complete inhibition. The Ca²⁺-channel activator S(–)BayK 8644 induced acidification when added in the dark and diminishedthe light-induced lowering of the extracellular pH. The Ca²⁺-ionophores,ionomycin and A23187 [GenBank] , also reduced the light response. Furthermore,the light-induced acidification was inhibited by the calmodulinantagonists W-7 and trifluoperazine, but not by W-5. These calmodulininhibitors completely inhibited the red light-induced acidification,but inhibited the response to blue light by only 60–70%.In general, inhibition by compounds affecting Ca-calmodulinsignalling was always stronger on the red light response thanthat on the blue light response (with the exception of verapamilthat blocked both the red and blue light responses equally well).This differential effect on red and blue light-induced responsesindicates a role for Ca²⁺-CaM signalling in both the red andblue light responses, while a second process, independent ofCa²⁺ is activated by blue light. Key words: Signal transduction, light-induced acidification, epidermal cells, pea     

Estimation of oxygen evolution by marine phytoplankton from measurement of the efficiency of Photosystem II electron flow

The relation between photosynthetic oxygen evolution and Photosystem II electron transport was investigated for the marine algae t Phaeodactylum tricornutum, Dunaliella tertiolecta, Tetraselmis sp., t Isochrysis sp. and t Rhodomonas sp.. The rate of Photosystem II electron transport was estimated from the incident photon flux density and the quantum efficiency of Photosystem II electron transport as determined by chlorophyll fluorescence. The relation between the estimated rate of Photosystem II electron transport and the rate of oxygen evolution was investigated by varying the ambient light intensity. At limiting light intensities a linear relation was found in all species. At intensities approaching light saturation, the relation was found to deviate from linearity. The slope of the line in the light-limited range is species dependent and related to differences in absorption cross-section of Photosystem II. The observed non-linearity at high irradiances is not caused by photorespiration but probably by a Mehler-type of oxygen reduction. The relationship could be modelled by including a redox-state dependent oxygen uptake. In the diatom t Phaeodactylum tricornutum, the photochemical efficiency of dark adapted open Photosystem II centers was found to be temperature-dependent with an optimum near 10°C.     

Effects of temperature and cycloheximide on secretion of cloned invertase from recombinant Saccharomyces cerevisiae

The effects of temperature on the kinetics and efficiency of secretion of cloned invertase were investigated in a recombinant yeast system. This system consisted of the baker's yeast Saccharomyces cerevisiae (SEY2102) transformed with the 2mu-based plasmid pBR58 which contains the entire SUC2 gene including the promoter, signal sequence, and structural gene. The recombinant yeast produces the naturally secreted yeast enzyme invertase. In transition experiments done at temperatures ranging from 25 degrees to 45 degrees C, the maximum invertase level and secretion rate exhibited maxima of 5.5 U/mL . OD and 4.6 U/mL . OD per hour, respectively, at 35 degrees C. Experiments involving the use of cycloheximide showed that it took approximately 15 min for secreted invertase to move through the secretion pathway, which held 0.4 U/mL . OD of specific activity. (c) 1995 John Wiley & Sons, Inc.     

Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy.

Specific molecular recognition events, detected by atomic force microscopy (AFM), so far lack the detailed topographical information that is usually observed in AFM. We have modified our AFM such that, in combination with a recently developed method to measure antibody-antigen recognition on the single molecular level (Hinterdorfer, P., W. Baumgartner, H. J. Gruber, K. Schilcher, and H. Schindler, Proc. Natl. Acad. Sci. USA 93:3477-3481 (1996)), it allows imaging of a submonolayer of intercellular adhesion molecule-1 (ICAM-1) in adhesion mode. We demonstrate that for the first time the resolution of the topographical image in adhesion mode is only limited by tip convolution and thus comparable to tapping mode images. This is demonstrated by imaging of individual ICAM-1 antigens in both the tapping mode and the adhesion mode. The contrast in the adhesion image that was measured simultaneously with the topography is caused by recognition between individual antibody-antigen pairs. By comparing the high-resolution height image with the adhesion image, it is possible to show that specific molecular recognition is highly correlated with topography. The stability of the improved microscope enabled imaging with forces as low as 100 pN and ultrafast scan speed of 22 force curves per second. The analysis of force curves showed that reproducible unbinding events on subsequent scan lines could be measured.     

Temperature-induced production of recombinant human insulin in high-cell density cultures of recombinant Escherichia coli

The construction of expression vectors encoding either the human insulin A- or B-chains fused to a synthetic peptide and the temperature-induced expression of the recombinant genes in Escherichia coli are reported. Using this two-chain approach we also describe the separate isolation of the insulin A- and B-chains from inclusion bodies and their subsequent assembly into native human insulin. The production of the insulin fusion proteins were carried out in high-cell density fed-batch cultures using a synthetic medium with glucose as sole carbon and energy source. The expression of the recombinant genes by temperature-shift in high-cell density cultures of recombinant E. coli resulted in product yields of grams per litre of culture broth, e.g. 4.5 g of insulin B-chain fusion protein per litre of culture broth. This translates into an expression yield of about 800 mg of the insulin B-chain per litre of culture. Under similar cultivation conditions the expression yield of the insulin A-chain corresponds to approximately 600 mg per litre of culture. The metabolic burden imposed on the recombinant cells during temperature-induced production of insulin fusion proteins in high-cell density cultures is reflected in an increased respiratory activity and a reduction of the biomass yield coefficient with respect to glucose.