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Mechanisms of the complex formation between plastocyanin and cytochrome f in higher plants (Spinacia oleracea and Brassica rapa), green microalgae Chlamydomonas reinhardtii and two species of cyanobacteria (Phormidium laminosum and Nostoc sp.) were investigated using combined Brownian and molecular dynamics simulations and hierarchical cluster analysis. In higher plants and green algae, electrostatic interactions force plastocyanin molecule close to the heme of cytochrome f. In the subsequent rotation of plastocyanin molecule around the point of electrostatic contact in the vicinity of cytochrome f, copper (Cu) atom approaches cytochrome heme forming a stable configuration where cytochrome f molecule behaves as a rather rigid body without conformational changes. In Nostoc plastocyanin molecule approaches cytochrome f in a different orientation (head‐on) where the stabilization of the plastocyanin–cytochrome f complex is accompanied by the conformational changes of the G188E189D190 loop that stabilizes the whole complex. In cyanobacterium P. laminosum, electrostatic preorientation of the approaching molecules was not detected, thus indicating that random motions rather than long‐range electrostatic interactions are responsible for the proper mutual orientation. We demonstrated that despite the structural similarity of the investigated electron transport proteins in different photosynthetic organisms, the complexity of molecular mechanisms of the complex formation increases in the following sequence: non‐heterocystous cyanobacteria – heterocystous cyanobacteria – green algae – flowering plants. [ABSTRACT FROM AUTHOR]

Pancreatic ductal adenocarcinoma (PDAC) represents the most common form of pancreatic cancer with rising incidence in developing countries and overall 5-year survival rates of less than 5%. The most frequent mutations in PDAC are gain-of-function mutations in KRAS as well as loss-of-function mutations in p53. Both mutations have severe impacts on the metabolism of tumor cells. Many of these metabolic changes are mediated by transporters or channels that regulate the exchange of metabolites and ions between the intracellular compartment and the tumor microenvironment. In the study presented here, our goal was to identify novel transporters or channels that regulate oxidative phosphorylation (OxPhos) in PDAC in order to characterize novel potential drug targets for the treatment of these cancers. We set up a Seahorse Analyzer XF based siRNA screen and identified previously described as well as novel regulators of OxPhos. The siRNA that resulted in the greatest change in cellular oxygen consumption was targeting the KCNN4 gene, which encodes for the Ca2+-sensitive K+ channel KCa3.1. This channel has not previously been reported to regulate OxPhos. Knock-down experiments as well as the use of a small molecule inhibitor confirmed its role in regulating oxygen consumption, ATP production and cellular proliferation. Furthermore, PDAC cell lines sensitive to KCa3.1 inhibition were shown to express the channel protein in the plasma membrane as well as in the mitochondria. These differences in the localization of KCa3.1 channels as well as differences in the regulation of cellular metabolism might offer opportunities for targeted therapy in subsets of PDAC. [ABSTRACT FROM AUTHOR]

Main conclusion: Components of the photosynthetic electron transport chain in pea (Pisum sativum L.) leaves under in vivo conditions showed the following sensitivity to the inhibitory action of chromium(VI): intersystem electron transport > photosystem I > photosystem II. Inhibitory effects of chromium (VI) (K2Cr2O7, Cr) on the light reactions of photosynthesis were studied in vivo in Pisum sativum L. by using Multi-function Plant Efficiency Analyser (M-PEA-2). Photosynthetic parameters related to photosystem (PS) II, PSI and intersystem electron carriers were calculated from the light-induced kinetics of prompt chlorophyll a fluorescence (OJIP transient), delayed chlorophyll a fluorescence (DF), and 820 nm modulated reflection (MR). We showed that the I2 step of DF induction is sensitive to inhibition of the Q0 site of the cytochrome b6f complex. Such parameters as δRo of the JIP test related to the functional state of photosynthetic reactions beyond the PQ pool, Vred of the MR induction assigned to the overall rate of P700+ and plastocyanin reduction, and I2 step of the DF induction were significantly altered in the presence of low-dose Cr(VI). Moderate doses of Cr affected mainly PSI-related parameters including Vox and ΔMR parameters of the MR induction, whereas high-dose treatment influenced JIP test parameters φPo(= FV/FM) and ψEo related to PSII. The obtained results showed that the earliest Cr(VI) effect on the photosynthetic electron transport chain manifests itself by inhibition of the intersystem electron transport, rather, at the level of the cytochrome b6f complex. Inhibitory effects of Cr on PSI were more pronounced than those on PSII. Sensitivity of the used kinetic parameters toward the functional state of photosynthetic reactions makes this approach suitable for early diagnostics of toxic action of pollutants on plants. [ABSTRACT FROM AUTHOR]