The extrinsic proteins of photosystem II in plants (PsbO, PsbP and PsbQ) are known to be targets of stress. In previous work, differential regulation of hypothetical isoforms of these proteins was observed in Nicotiana benthamiana upon viral infection. Each of these proteins is encoded by a multigene family in this species: there are at least four genes encoding PsbO and PsbP and two encoding PsbQ. The results of structural and functional analyses suggest that PsbO and PsbP isoforms could show differences in activity, based on significant substitutions in their primary structure. Two psbQ sequences were isolated which encode identical mature proteins. and M. I. Pérez-Bueno ... [et al.].
PsbP is an extrinsic protein of PSII having a function of Ca2+ and Cl- retention in the water-oxidizing center (WOC). In order to understand the mechanism how PsbP regulates the Cl- binding in WOC, we examined the effect of PsbP depletion on the protein structures around the Cl- sites using Fourier transform infrared (FTIR) spectroscopy. Light-induced FTIR difference spectra upon the S1-S2 transition were obtained using Cl--bound and NO3--substituted PSII membranes in the presence and absence of PsbP. A clear difference in the amide I band changes by PsbP depletion was observed between Cl--bound and NO3--substituted PSII samples, indicating that PsbP binding perturbed the protein conformations around the Cl-ion(s) in WOC. It is suggested that PsbP stabilizes the Cl- binding by regulating the dissociation constant of Cl- and/or an energy barrier of Cl- dissociation through protein conformational changes around the Cl- ion(s)., J. Kondo, T. Noguchi., and Obsahuje bibliografické odkazy
The PsbM (3.9 kDa) and PsbY (4.2 kDa) proteins are membrane-spanning, single-helix, subunits associated with the chlorophyll-binding CP47 pre-complex of photosystem II (PSII). Removal of PsbM resulted in accumulation of PSII pre-assembly complexes and impaired electron transfer between the primary (QA) and secondary (QB) plastoquinone electron acceptors of PSII indicating that the QB-binding site and bicarbonate binding to the non-heme iron were altered in this strain. Removal of PsbY alone had only a minor impact on PSII activity but deleting PsbY in the PsbM background led to additional modification of the acceptor side resulting in PsbM:PsbY cells being susceptible to photodamage and this required protein synthesis for recovery. Addition of bicarbonate was able to compensate for the light-induced damage in PsbM:PsbY cells potentially re-occupying the modified bicarbonate-binding site in the PsbM:PsbY strain and complementation of PsbM:PsbY cells with the psbY gene restored the PsbM phenotype., S. Biswas, J. J. Eaton-Rye., and Obsahuje bibliografické odkazy
In this study, the JIP test was used to assess the drought tolerance of two sweet cherry cultivars (Prunus avium L.) (modern and autochthonous). Plants were exposed to progressive drought by withholding water and their fast (< 1 s) chlorophyll fluorescence kinetics was evaluated. JIP test analysis showed that drought stress caused a greater decrease in performance indices (PIABS and PItotal) in a modern cultivar, as compared to an autochthonous one. Our results suggest that limited reoxidation of primary quinone electron acceptor (QA), higher amount of secondary quinone electron acceptor (QB-) nonreducing reaction centres, or inhibition of the electron transport between QA and QB, decreased more seriously the photosynthetic performance of the modern cultivar. Further, higher positive L- and K-bands observed for the modern cultivar also suggest lower energetic connectivity between PSII units and increased inhibition of oxygen-evolving complex over autochthonous cultivar. Our results suggest that the autochthonous cultivar Crveni hrušt had better photosynthetic performance under drought conditions, compared to the modern cultivar New Star.
The cytosolic and chloroplastic isoforms of glucose-6-phosphate dehydrogenase (G6PDH) were separated and purified from barley leaves (Hordeum vulgare L.). In etiolated leaves, only the cytosolic isoform was expressed. The molecular mass of the cytosolic enzyme, G6PDH1, was 112±8 kDa and that of the chloroplast enzyme, G6PDH2, was 136±7 kDa. The Km values for glucose-6-phosphate and NADP were 0.133 and 0.041 mM for G6PDH1, and 0.275 and 0.062 mM for G6PDH2, respectively. The pH optimum was 8.2 for G6PDH1 and 7.8 for G6PDH2. The enzyme is absolutely specific for NADP. NADPH is a competitive inhibitor of the G6PDH1 in respect to glucose-6-phosphate (G6P) and NADP (Ki = 0.050 and 0.025 mM, respectively). NADPH is a competitive inhibitor of the G6PDH2 in respect to NADP (Ki = 0.010 mM), but a non-competitive inhibitor in respect to the G6P. ADP, AMP, UTP, NAD, and NADH had no effect on the activity of G6PDH. ATP inhibited the G6PDH2 activity. and A. Semenihina ... [et al.].
A traditional method is reported for purification of phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) from leaves of Amaranthus hypochondriacus L. with a high yield of 50 %, 135-fold purification, and specific activity of 900 mmol kg-1(protein) s-1. PEPC was purified from light-adapted leaves of A. hypochondriacus, involving 40-60 % ammonium sulphate fractionation, followed by chromatography on columns of DEAE-Sepharose, hydroxylapatite (HAP), and Seralose 6-B. The enzyme appeared as a single band on 10 % SDS-PAGE, with a molecular mass of about 100 kDa. Kinetic studies with purified enzyme confirmed the PEPC to be the light-form of the enzyme. Glycerol generally increased the stability of PEPC. The stability and storage of the purified enzyme was studied at temperatures of 4 °C, -20 °C, and liquid nitrogen. PEPC maintained its activity for up to 3 months upon storage with 50 % (v/v) glycerol in liquid nitrogen. and J. Gayathri, K. Parvathi, A. S. Raghavendra.
Etiolated oat mesophyll protoplasts and etioplasts released from etiolated protoplasts were able to perform protochlorophyliide (PChlide) reduction but showed a high amount of inactive P633/628» compared with intact leaves. In špite of this, the anabolic reduction charge (ARC, NAPDH+H+/NADP+ + NADPF1+H+) was maintained at 0.66 - 0.77 up to 30 min of irradiation with weak "white light" (10 W m‘2). This indicated a high buffering capacity of the isolated systém to provide NADPF1+H+, only at the expense of intemal reserves. A change in the redox statě of the NADP+/NADPH+H^ couple during protochlorophyliide reduction was not observed in etioplasts. The resulting redox potential was in the range of -330 mV optimal for protochlorophyliide reduction.
Binding kinetics of some photosystem 2 (PS 2)-directed ínhibitors to isolated pea thylakoids during photoinUbition were studied. Changes in the herbicide binding activity were overshadowed by the redox statě of the plastoquinone (PQ) pool. After extraction of the PQ pool from the thylakoids, a correlation between the decline of herbicide binding activity and the electron transfer activity during strong irradiation was found in the presence of oxygen; in anaerobiosis this correlation was absent. The opposite was observed for the binding kinetics of the electron transfer acceptor 2,6-dichlorophenol indophenol, that was affected faster under anaerobic conditions. Following irradiation, a movement of damaged Dj-protein products from the appressed to the non-appressed regions of the membrane was observed. This observation was correlated to the fínding of an increased level of atrazine bound to the non-appressed regions of irradiated membranes.