BBA - Bioenergetics (v.1797, #5)
Editorial Board (i).
A revised energy partitioning approach to assess the yields of non-photochemical quenching components by C.R. Guadagno; A. Virzo De Santo; N. D'Ambrosio (525-530).
Non-photochemical quenching (NPQ) is a complex and still unclear mechanism essential for higher plants. The intensive research on this subject has highlighted three main components of NPQ: energy-dependent process (qE); state transitions to balance the excitation of PSII and PSI (qT); and photoinhibitory processes (qI). Recently, these components have been resolved as quantum yields according to the energy partitioning approach that takes into account the rate constants of every process involved in the quenching mechanisms of excited chlorophylls. In this study a fully extended quantum yield approach and the introduction of novel equations to assess the yields of each NPQ component are presented. Furthermore, a complete analysis of the yield of NPQ in Beta vulgaris exposed to different irradiances has been carried out. In agreement with experimental results here it is shown that the previous approach may amplify the yield of qE component and flatten the quantitative results of fluorescence analysis. Moreover, the significance of taking into account the physiological variability of NPQ for a correct assessment of energy partitioning is demonstrated.
Keywords: Chlorophyll a fluorescence; Quantum yield convention; NPQ; Thermal dissipation; Beta vulgaris;
Horseradish peroxidase-catalyzed oxidation of chlorophyll a with hydrogen peroxide by Paavo H. Hynninen; Vesa Kaartinen; Erkki Kolehmainen (531-542).
Horseradish peroxidase was verified to catalyze, without any phenol, the hydrogen peroxide oxidation of chlorophyll a (Chl a), solubilized with Triton X-100. The 132(S) and 132(R) diastereomers of 132-hydroxyChl a were characterized as major oxidation products (ca. 60%) by TLC on sucrose, UV–vis, 1H, and 13C NMR spectra, as well as fast-atom bombardment MS. A minor amount of the 152-methyl, 173-phytyl ester of Mg-unstable chlorin was identified on the basis of its UV–vis spectrum and reactivity with diazomethane, which converted it to the 131,152-dimethyl, 173-phytyl ester of Mg-purpurin 7. The side products (ca. 10%) were suggested to include the 173-phytyl ester of Mg-purpurin 18, which is known to form easily from the Mg-unstable chlorin. The side products also included two red components with UV–vis spectral features resembling those of pure Chl a enolate anion. Hence, the two red components were assigned to the enolate anions of Chl a and pheophytin a or, alternatively, two different complexes of the Chl a enolate ion with Triton X-100. All the above products characterized by us are included in our published free-radical allomerization mechanism of Chl a, i.e. oxidation by ground-state dioxygen. The HRP clearly accelerated the allomerization process, but it did not produce bilins, that is, open-chain tetrapyrroles, the formation of which would require oxygenolysis of the chlorin macrocycle. In this regard, our results are in discrepancy with the claim by several researchers that ‘bilirubin-like compounds’ are formed in the HRP-catalyzed oxidation of Chl a. Inspection of the likely reactions that occurred on the distal side of the heme in the active centre of HRP provided a reasonable explanation for the observed catalytic effect of the HRP on the allomerization of Chl. In the active centre of HRP, the imidazole nitrogen of His-42 was considered to play a crucial role in the C-132 deprotonation of Chl a, which resulted in the Chl a enolate ion resonance hybrid. The Chl enolate was then oxidized to the Chl 132-radical while the HRP Compound I was reduced to Compound II. The same reactive Chl derivatives, i.e. the Chl enolate anion and the Chl 132-radical, which are produced twice in the HRP reaction cycle, happen to be the crucial intermediates in the initial stages of the Chl allomerization mechanism.
Keywords: Enzyme; Peroxidase; Chlorophyll; Allomerization; Oxidation; Free-radical;
Oligomerization and pigmentation dependent excitation energy transfer in fucoxanthin–chlorophyll proteins by Nina Gildenhoff; Sergiu Amarie; Kathi Gundermann; Anja Beer; Claudia Büchel; Josef Wachtveitl (543-549).
The ultrafast caroteonid to chlorophyll a energy transfer dynamics of the isolated fucoxanthin–chlorophyll proteins FCPa and FCPb from the diatom Cyclotella meneghiniana was investigated in a comprehensive study using transient absorption in the visible and near infrared spectral region as well as static fluorescence spectroscopy. The altered oligomerization state of both antenna systems results in a more efficient energy transfer for FCPa, which is also reflected in the different chlorophyll a fluorescence quantum yields. We therefore assume an increased quenching in the higher oligomers of FCPb. The influence of the carotenoid composition was investigated using FCPa and FCPb samples grown under different light conditions and excitation wavelengths at the blue (500 nm) and red (550 nm) wings of the carotenoid absorption. The different light conditions yield in altered amounts of the xanthophyll cycle pigments diadinoxanthin and diatoxanthin. Since no significant dynamic changes are observed for high light and low light samples, the contribution of the xanthophyll cycle pigments to the energy transfer is most likely negligible. On the contrary, the observed dynamics change drastically for the different excitation wavelengths. The analyses of the decay associated spectra of FCPb suggest an altered energy transfer pathway. For FCPa even an additional time constant was found after excitation at 500 nm. It is assigned to the intrinsic lifetime of either the xanthophyll cycle carotenoids or more probable the blue absorbing fucoxanthins. Based on our studies we propose a detailed model explaining the different excitation energy transfer pathways in FCPa.
Keywords: Fucoxanthin–chlorophyll protein; Ultrafast spectroscopy; Cyclotella meneghiniana; Excitation energy transfer; Light harvesting complex; Diatom;
A pathogenic mutation in cytochrome c oxidase results in impaired proton pumping while retaining O2-reduction activity by Ida Namslauer; Hyun Ju Lee; Robert B. Gennis; Peter Brzezinski (550-556).
In this work we have investigated the effect of a pathogenic mitochondrial DNA mutation found in human colon cells, at a functional-molecular level. The mutation results in the amino-acid substitution Tyr19His in subunit I of the human CytcO and it is associated with respiratory deficiency. It was introduced into Rhodobacter sphaeroides, which carries a cytochrome c oxidase (cytochrome aa 3) that serves as a model of the mitochondrial counterpart. The residue is situated in the middle of a pathway that is used to transfer substrate protons as well as protons that are pumped across the membrane. The Tyr33His (equivalent residue in the bacterial CytcO) structural variant of the enzyme was purified and its function was investigated. The results show that in the structurally altered CytcO the activity decreased due to slowed proton transfer; proton transfer from an internal proton donor, the highly-conserved Glu286, to the catalytic site was slowed by a factor of ∼ 5, while reprotonation of the Glu from solution was slowed by a factor of ∼ 40. In addition, in the structural variant proton pumping was completely impaired. These results are explained in terms of introduction of a barrier for proton transfer through the D pathway and changes in the coordination of water molecules surrounding the Glu286 residue. The study offers an explanation, at the molecular level, to the link between a specific amino-acid substitution and a pathogenic phenotype identified in human colon cells.
Keywords: Respiratory chain; Electron transfer; Mitochondrial disease; Cytochrome aa 3; Mitochondria; Deficiency; mtDNA;
Kinetic model of the inhibition of respiration by endogenous nitric oxide in intact cells by Enara Aguirre; Félix Rodríguez-Juárez; Andrea Bellelli; Erich Gnaiger; Susana Cadenas (557-565).
Nitric oxide (NO) inhibits mitochondrial respiration by decreasing the apparent affinity of cytochrome c oxidase (CcO) for oxygen. Using iNOS-transfected HEK 293 cells to achieve regulated intracellular NO production, we determined NO and O2 concentrations and mitochondrial O2 consumption by high-resolution respirometry over a range of O2 concentrations down to nanomolar. Inhibition of respiration by NO was reversible, and complete NO removal recovered cell respiration above its routine reference values. Respiration was observed even at high NO concentrations, and the dependence of IC50 on [O2] exhibits a characteristic but puzzling parabolic shape; both these features imply that CcO is protected from complete inactivation by NO and are likely to be physiologically relevant. We present a kinetic model of CcO inhibition by NO that efficiently predicts experimentally determined respiration at physiological O2 and NO concentrations and under hypoxia, and accurately predicts the respiratory responses under hyperoxia. The model invokes competitive and uncompetitive inhibition by binding of NO to the reduced and oxidized forms of CcO, respectively, and suggests that dissociation of NO from reduced CcO may involve its O2-dependent oxidation. It also explains the non-linear dependence of IC50 on O2 concentration, and the hyperbolic increase of c 50 as a function of NO concentration.
Keywords: Nitric oxide; Mitochondrial respiration; Cytochrome c oxidase; Oxygen consumption; Mitochondria; Kinetic model;
Role of FtsH2 in the repair of Photosystem II in mutants of the cyanobacterium Synechocystis PCC 6803 with impaired assembly or stability of the CaMn4 cluster by Josef Komenda; Jana Knoppová; Vendula Krynická; Peter J. Nixon; Martin Tichý (566-575).
The FtsH2 protease, encoded by the slr0228 gene, plays a key role in the selective degradation of photodamaged D1 protein during the repair of Photosystem II (PSII) in the cyanobacterium Synechocystis sp. PCC 6803. To test whether additional proteases might be involved in D1 degradation during high rates of photodamage, we have studied the synthesis and degradation of the D1 protein in ΔPsbO and ΔPsbV mutants, in which the CaMn4 cluster catalyzing oxygen evolution is less stable, and in the D1 processing mutants, D1-S345P and ΔCtpA, which are unable to assemble a functional cluster. All four mutants exhibited a dramatically increased rate of D1 degradation in high light compared to the wild-type. Additional inactivation of the ftsH2 gene slowed the rate of D1 degradation dramatically and increased the level of PSII complexes. We conclude that FtsH2 plays a major role in the degradation of both precursor and mature forms of D1 following donor-side photoinhibition. However, this conclusion concerned only D1 assembled into larger complexes containing at least D2 and CP47. In the ΔpsbEFLJ deletion mutant blocked at an early stage in PSII assembly, unassembled D1 protein was efficiently degraded in the absence of FtsH2 pointing to the involvement of other protease(s). Significantly, the ΔPsbO mutant displayed unusually low levels of cellular chlorophyll at extremely low-light intensities. The possibilities that PSII repair may limit the availability of chlorophyll for the biogenesis of other chlorophyll-binding proteins and that PsbO might have a regulatory role in PSII repair are discussed.
Keywords: CtpA protease; D1 degradation and maturation; FtsH protease; Photosystem II; psbA gene; psbO gene; psbV gene; Synechocystis PCC 6803;
Erratum to “Purification characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis” [Biochim. Biophys. Acta 1797 (2010) 160–166] by Ryo Nagao; Tatsuya Tomo; Eri Noguchi; Saori Nakajima; Takehiro Suzuki; Akinori Okumura; Yasuhiro Kashino; Mamoru Mimuro; Masahiko Ikeuchi; Isao Enami (576).