BBA - Bioenergetics (v.1858, #6)
Tyrozine D oxidation and redox equilibrium in photosystem II by Nigar Ahmadova; Felix M. Ho; Stenbjörn Styring; Fikret Mamedov (407-417).
Tyrosine D (TyrD) is an auxiliary redox active tyrosine residue in photosystem II (PSII). The mechanism of TyrD oxidation was investigated by EPR spectroscopy, flash-induced fluorescence decay and thermoluminescence measurements in PSII enriched membranes from spinach. PSII membranes were chemically treated with 3 mM ascorbate and 1 mM diaminodurene and subsequent washing, leading to the complete reduction of TyrD. TyrD oxidation kinetics and competing recombination reactions were measured after a single saturating flash in the absence and presence of DCMU (inhibitor of the QB-site) in the pH range of 4.7–8.5. Two kinetic phases of TyrD oxidation were observed by the time resolved EPR spectroscopy – the fast phase (msec-sec time range) and the pH dependent slow phase (tens of seconds time range). In the presence of DCMU, TyrD oxidation kinetics was monophasic in the entire pH range, i.e. only the fast kinetics was observed. The results obtained from the fluorescence and thermoluminescence analysis show that when forward electron transport is blocked in the presence of DCMU, the S2QA − recombination outcompetes the slow phase of TyrD oxidation by the S2 state. Modelling of the whole complex of these electron transfer events associated with TyrD oxidation fitted very well with our experimental data. Based on these data, structural information and theoretical considerations we confirm our assignment of the fast and slow oxidation kinetics to two populations of PSII centers with different water positions (proximal and distal) in the TyrD vicinity.
Keywords: Photosystem II; Electron transfer; QA − S2 state recombination; Tyrosine D;
Synchronism in mitochondrial ROS flashes, membrane depolarization and calcium sparks in human carcinoma cells by Andrey V. Kuznetsov; Sabzali Javadov; Valdur Saks; Raimund Margreiter; Michael Grimm (418-431).
Mitochondria are major producers of reactive oxygen species (ROS) in many cells including cancer cells. However, complex interrelationships between mitochondrial ROS (mitoROS), mitochondrial membrane potential (ΔΨm) and Ca2 + are not completely understood. Using human carcinoma cells, we further highlight biphasic ROS dynamics: - gradual mitoROS increase followed by mitoROS flash. Also, we demonstrate heterogeneity in rates of mitoROS generation and flash initiation time. Comparing mitochondrial and near-extra-mitochondrial signals, we show that mechanisms of mitoROS flashes in single mitochondria, linked to mitochondrial permeability transition pore opening (ΔΨm collapse) and calcium sparks, may involve flash triggering by certain levels of external ROS released from the same mitochondria. In addition, mitochondria-mitochondria interactions can produce wave propagations of mitoROS flashes and ΔΨm collapses in cancer cells similar to phenomena of ROS-induced ROS release (RIRR). Our data suggest that in cancer cells RIRR, activation of mitoROS flashes and mitochondrial depolarization may involve participation of extramitochondrial-ROS produced either by individual mitochondria and/or by neighboring mitochondria. This could represent general mechanisms in ROS-ROS signaling with suggested role in both mitochondrial and cellular physiology and signaling.
Keywords: Carcinoma cells; Ca2 + sparks; Mitochondria; Membrane potential; ROS flashes;
Charged groups at binding interfaces of the PsbO subunit of photosystem II: A combined bioinformatics and simulation study by Coral del Val; Ana-Nicoleta Bondar (432-441).
PsbO is an extrinsic subunit of photosystem II engaged in complex binding interactions within photosystem II. At the interface between PsbO, D1 and D2 subunits of photosystem II, a cluster of charged and polar groups of PsbO is part of an extended hydrogen-bond network thought to participate in proton transfer. The precise role of specific amino acid residues at this complex binding interface remains a key open question. Here, we address this question by carrying out extensive bioinformatics analyses and molecular dynamics simulations of PsbO proteins with mutations at the binding interface. We find that PsbO proteins from cyanobacteria vs. plants have specific preferences for the number and composition of charged amino acid residues that may ensure that PsbO proteins avoid aggregation and expose long unstructured loops for binding to photosystem II. A cluster of conserved charged groups with dynamic hydrogen bonds provides PsbO with structural plasticity at the binding interface with photosystem II.
Identity and function of a cardiac mitochondrial small conductance Ca2+-activated K+ channel splice variant by MeiYing Yang; Amadou K.S. Camara; Mohammed Aldakkak; Wai-Meng Kwok; David F. Stowe (442-458).
We provide evidence for location and function of a small conductance, Ca2+-activated K+ (SKCa) channel isoform 3 (SK3) in mitochondria (m) of guinea pig, rat and human ventricular myocytes. SKCa agonists protected isolated hearts and mitochondria against ischemia/reperfusion (IR) injury; SKCa antagonists worsened IR injury. Intravenous infusion of a SKCa channel agonist/antagonist, respectively, in intact rats was effective in reducing/enhancing regional infarct size induced by coronary artery occlusion. Localization of SK3 in mitochondria was evidenced by Western blot of inner mitochondrial membrane, immunocytochemical staining of cardiomyocytes, and immunogold labeling of isolated mitochondria. We identified a SK3 splice variant in guinea pig (SK3.1, aka SK3a) and human ventricular cells (SK3.2) by amplifying mRNA, and show mitochondrial expression in mouse atrial tumor cells (HL-1) by transfection with full length and truncated SK3.1 protein. We found that the N-terminus is not required for mitochondrial trafficking but the C-terminus beyond the Ca2+ calmodulin binding domain is required for Ca2+ sensing to induce mK+ influx and/or promote mitochondrial localization. In isolated guinea pig mitochondria and in SK3 overexpressed HL-1 cells, mK+ influx was driven by adding CaCl2. Moreover, there was a greater fall in membrane potential (ΔΨm), and enhanced cell death with simulated cell injury after silencing SK3.1 with siRNA. Although SKCa channel opening protects the heart and mitochondria against IR injury, the mechanism for favorable bioenergetics effects resulting from SKCa channel opening remains unclear. SKCa channels could play an essential role in restraining cardiac mitochondria from inducing oxidative stress-induced injury resulting from mCa2+ overload.
Keywords: Cardiac mitochondria; Inner mitochondrial membrane; Cell signaling; Ischemia reperfusion injury; Oxidant stress; Small conductance Ca2+-sensitive K+ channel;
Connectivity among Photosystem II centers in phytoplankters: Patterns and responses by Kui Xu; Jessica L. Grant-Burt; Natalie Donaher; Douglas A. Campbell (459-474).
Fast Repetition and Relaxation chlorophyll fluorescence induction is used to estimate the effective absorption cross section of PSII (σPSII), to analyze phytoplankton acclimation and electron transport. The fitting coefficient ρ measures excitation transfer from closed PSII to remaining open PSII upon illumination, which could theoretically generate a progressive increase in σPSII for the remaining open PSII. To investigate how ρ responds to illumination we grew marine phytoplankters with diverse antenna structures (Prochlorococcus, Synechococcus, Ostreococcus and Thalassiosira pseudonana) under limiting or saturating growth light. Initial ρ varied with growth light in Synechococcus and Thalassiosira. With increasing actinic illumination PSII closed progressively and ρ decreased for all four taxa, in a pattern explicable as an exponential decay of ρ with increasing distance between remaining open PSII reaction centers. This light-dependent down-regulation of ρ allows the four phytoplankters to limit the effect of increasing light upon σPSII. The four structurally distinct taxa showed, however, distinct rates of response of ρ to PSII closure, likely reflecting differences in the spacing or orientation among their PSII centers. Following saturating illumination recovery of ρ in darkness coincided directly with PSII re-opening in Prochlorococcus. Even after PSII had re-opened in Synechococcus a transition to State II slowed dark recovery of ρ. In Ostreococcus sustained NPQ slowed dark recovery of ρ. In Thalassiosira dark recovery of ρ was slowed, possibly by a light-induced change in PSII spacing. These patterns of ρ versus PSII closure are thus a convenient probe of comparative PSII spacings.
Keywords: Fast Repetition and Relaxation chlorophyll fluorescence induction; Light acclimation; Ostreococcus; Photosynthesis; Phytoplankton; Prochlorococcus; Synechococcus; Thalassiosira pseudonana;