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1.
  • Alam, Muneeba Zubair, et al. (author)
  • Soluble and hydrolyzable phenolic compounds in date fruits (Phoenix dactylifera L.) by UPLC-QTOF-MS/MS and UPLC-DAD
  • 2024
  • In: Journal of Food Composition and Analysis. - 0889-1575 .- 1096-0481. ; 132
  • Journal article (peer-reviewed)abstract
    • Phenolic compounds in plant foods exist in soluble, hydrolyzable, and insoluble forms. This study is the first to analyze both soluble (extracted using 80 % methanol) and hydrolyzable (extracted using ethyl acetate after alkaline hydrolysis) phenolics in 18 date cultivars. The phenolic compounds were identified and quantified using ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry and diode array detectors. A total of 59 peaks were detected, of which 45 were identified while 14 remained unidentified. The concentrations of the soluble and hydrolyzable phenolics ranged from 27.5–54.0 and 24–78.5 mg/100 g fresh weight, respectively. Gallic acid and ferulic acid predominated in the soluble and hydrolyzable fractions, quercetin and taxifolin were predominant in the soluble fraction, while caffeic acid and chlorogenic acid were abundant in the hydrolyzable fraction. The hydrolyzable phenolics were found to contribute significantly to the total phenolic content of date fruits pointing to the need to include them in the analysis of total phenolic compounds in foods.
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2.
  • Cheng, Ken, 1987, et al. (author)
  • An LC-QToF MS based method for untargeted metabolomics of human fecal samples
  • 2020
  • In: Metabolomics. - : Springer Science and Business Media LLC. - 1573-3882 .- 1573-3890. ; 16:4
  • Journal article (peer-reviewed)abstract
    • Introduction: Consensus in sample preparation for untargeted human fecal metabolomics is lacking. Objectives: To obtain sample preparation with broad metabolite coverage for high-throughput LC–MS. Methods: Extraction solvent, solvent ratio and fresh frozen-vs-lyophilized samples were evaluated by metabolite feature quality. Results: Methanol at 5 mL per g wet feces provided a wide metabolite coverage with optimal balance between signal intensity and saturation for both fresh frozen and lyophilized samples. Lyophilization did not affect SCFA and is recommended because of convenience in normalizing to dry matter. Conclusion: The suggested sample preparation is simple, efficient and suitable for large-scale human fecal metabolomics.
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3.
  • Eriksen, Anne Kirstine, et al. (author)
  • Effects of whole-grain wheat, rye, and lignan supplementation on cardiometabolic risk factors in men with metabolic syndrome: A randomized crossover trial
  • 2020
  • In: American Journal of Clinical Nutrition. - : Elsevier BV. - 0002-9165 .- 1938-3207. ; 111:4, s. 864-876
  • Journal article (peer-reviewed)abstract
    • A whole-grain (WG)-rich diet has shown to have potential for both prevention and treatment of the metabolic syndrome (MetS), which is a cluster of risk factors that increase the risk of type 2 diabetes and cardiovascular disease. Different WGs may have different health effects. WG rye, in particular, may improve glucose homeostasis and blood lipids, possibly mediated through fermentable dietary fiber and lignans. Recent studies have also suggested a crucial role of the gut microbiota in response to WG. Objectives: The aim was to investigate WG rye, alone and with lignan supplements [secoisolariciresinol diglucoside (SDG)], and WG wheat diets on glucose tolerance [oral-glucose-tolerance test (OGTT)], other cardiometabolic outcomes, enterolignans, and microbiota composition. Moreover, we exploratively evaluated the role of gut microbiota enterotypes in response to intervention diets. Methods: Forty men with MetS risk profile were randomly assigned to WG diets in an 8-wk crossover study. The rye diet was supplemented with 280 mg SDG at weeks 4-8. Effects of treatment were evaluated by mixed-effects modeling, and effects on microbiota composition and the role of gut microbiota as a predictor of response to treatment were analyzed by random forest plots. Results: The WG rye diet (± SDG supplements) did not affect the OGTT compared with WG wheat. Total and LDL cholesterol were lowered (-0.06 and -0.09 mmol/L, respectively; P < 0.05) after WG rye compared with WG wheat after 4 wk but not after 8 wk. WG rye resulted in higher abundance of Bifidobacterium [fold-change (FC) = 2.58, P < 0.001] compared with baseline and lower abundance of Clostridium genus compared with WG wheat (FC = 0.54, P = 0.02). The explorative analyses suggest that baseline enterotype is associated with total and LDL-cholesterol response to diet. Conclusions: WG rye, alone or with SDG supplementation, compared with WG wheat did not affect glucose metabolism but caused transient LDL-cholesterol reduction. The effect of WG diets appeared to differ according to enterotype. This trial was registered at www.clinicaltrials.gov as NCT02987595.
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4.
  • Fristedt, Rikard, 1978, et al. (author)
  • Quantitation of circulating short-chain fatty acids in small volume blood samples from animals and humans
  • 2024
  • In: Talanta. - 0039-9140. ; 272
  • Journal article (peer-reviewed)abstract
    • Background: The role of gut microbiota in human health has been intensively studied and more recently shifted from emphasis on composition towards function. Function is partly mediated through formed metabolites. Short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate as well as their branched analogues represent major products from gut fermentation of dietary fibre and proteins, respectively. Robust and high-throughput analysis of SCFAs in small volume blood samples have proven difficult. Major obstacles come from the ubiquitous presence of SCFAs that leads to contaminations and unstable analytical results because of the high volatility of these small molecules. Comprehensive and comparable data on the variation of SCFAs in blood samples from different blood matrices and mammal species including humans is lacking. Therefore, our aim was to develop and evaluate a stable and robust method for quantitation of 8 SCFAs and related fermentation products in small volume blood plasma samples and to investigate their variation in humans and different animal species. Results: Derivatization was a successful approach for measurement of SCFAs in biological samples but quenching of the derivatization reaction was crucial to obtain long-term stability of the derivatized analytes. In total 9 compounds (including succinic acid) were separated in 5 min. The method was linear over the range 0.6–3200 nM formic (FA), acetic (AA), 0.3–1600 nM propionic (PA), and 0.16–800 nM for butyric (BA)-, isobutyric (IBA)-, valeric (VA)-, isovaleric (IVA)-, succinic (SA) and caproic acid (CA). The precision ranged ≤12 % within days and ≤28 % between days (except for CA and VA) in three different plasma quality control (QC) samples (29 batches analyzed over 3 months). The extraction recovery was on average 94 % for the different SCFAs. Typical interquartile range (IQR) concentrations (μM) of SCFAs in human plasma samples were 168 μM (FA), 64 μM (AA), 2.2 μM (PA), 0.54 μM (BA), 0.66 μM (IBA), 0.18 μM (VA), 0.40 μM (IVA), and 0.34 μM (CA). In total, 55 samples per batch/day were successfully analyzed and in total 5380 human plasma samples measured over a 3-year timespan. Significance: The developed UHPLC-MS based method was suitable for measuring SCFAs in small blood volume samples and enabled robust quantitative data.
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5.
  • Iversen, Kia Noehr, 1987, et al. (author)
  • The Effects of High Fiber Rye, Compared to Refined Wheat, on Gut Microbiota Composition, Plasma Short Chain Fatty Acids, and Implications for Weight Loss and Metabolic Risk Factors (the RyeWeight Study)
  • 2022
  • In: Nutrients. - : MDPI AG. - 2072-6643 .- 2072-6643. ; 14:8
  • Journal article (peer-reviewed)abstract
    • Consumption of whole grain and cereal fiber have been inversely associated with body weight and obesity measures in observational studies but data from large, long-term randomized interventions are scarce. Among the cereals, rye has the highest fiber content and high rye consumption has been linked to increased production of gut fermentation products, as well as reduced risks of obesity and metabolic disease. The effects on body weight and metabolic risk factors may partly be mediated through gut microbiota and/or their fermentation products. We used data from a randomized controlled weight loss trial where participants were randomized to a hypocaloric diet rich in either high fiber rye foods or refined wheat foods for 12 weeks to investigate the effects of the intervention on gut microbiota composition and plasma short chain fatty acids, as well as the potential association with weight loss and metabolic risk markers. Rye, compared to wheat, induced some changes in gut microbiota composition, including increased abundance of the butyrate producing Agathobacter and reduced abundance of [Ruminococcus] torques group, which may be related to reductions in low grade inflammation caused by the intervention. Plasma butyrate increased in the rye group. In conclusion, intervention with high fiber rye foods induced some changes in gut microbiota composition and plasma short chain fatty acid concentration, which were associated with improvements in metabolic risk markers as a result of the intervention.
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6.
  • Jernfors, Toni, et al. (author)
  • Association between gut health and gut microbiota in a polluted environment
  • 2024
  • In: Science of the Total Environment. - 0048-9697 .- 1879-1026. ; 914
  • Journal article (peer-reviewed)abstract
    • Animals host complex bacterial communities in their gastrointestinal tracts, with which they share a mutualistic interaction. The numerous effects these interactions grant to the host include regulation of the immune system, defense against pathogen invasion, digestion of otherwise undigestible foodstuffs, and impacts on host behaviour. Exposure to stressors, such as environmental pollution, parasites, and/or predators, can alter the composition of the gut microbiome, potentially affecting host-microbiome interactions that can be manifest in the host as, for example, metabolic dysfunction or inflammation. However, whether a change in gut microbiota in wild animals associates with a change in host condition is seldom examined. Thus, we quantified whether wild bank voles inhabiting a polluted environment, areas where there are environmental radionuclides, exhibited a change in gut microbiota (using 16S amplicon sequencing) and concomitant change in host health using a combined approach of transcriptomics, histological staining analyses of colon tissue, and quantification of short-chain fatty acids in faeces and blood. Concomitant with a change in gut microbiota in animals inhabiting contaminated areas, we found evidence of poor gut health in the host, such as hypotrophy of goblet cells and likely weakened mucus layer and related changes in Clca1 and Agr2 gene expression, but no visible inflammation in colon tissue. Through this case study we show that inhabiting a polluted environment can have wide reaching effects on the gut health of affected animals, and that gut health and other host health parameters should be examined together with gut microbiota in ecotoxicological studies.
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7.
  • Verspoor, Rudi L., et al. (author)
  • Mineral analysis reveals extreme manganese concentrations in wild harvested and commercially available edible termites
  • 2020
  • In: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322 .- 2045-2322. ; 10:1
  • Journal article (peer-reviewed)abstract
    • Termites are widely used as a food resource, particularly in Africa and Asia. Markets for insects as food are also expanding worldwide. To inform the development of insect-based foods, we analysed selected minerals (Fe-Mn-Zn-Cu-Mg) in wild-harvested and commercially available termites. Mineral values were compared to selected commercially available insects. Alate termites, of the genera Macrotermes and Odontotermes, showed remarkably high manganese (Mn) content (292-515 mg/100 gdw), roughly 50-100 times the concentrations detected in other insects. Other mineral elements occur at moderate concentrations in all insects examined. On further examination, the Mn is located primarily in the abdomens of the Macrotermes subhyalinus; with scanning electron microscopy revealing small spherical structures highly enriched for Mn. We identify the fungus comb, of Macrotermes subhyanus, as a potential biological source of the high Mn concentrations. Consuming even small quantities of termite alates could exceed current upper recommended intakes for Mn in both adults and children. Given the widespread use of termites as food, a better understanding the sources, distribution and bio-availability of these high Mn concentrations in termite alates is needed.
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8.
  • Amstutz, Cynthia L., et al. (author)
  • An atypical short-chain dehydrogenase–reductase functions in the relaxation of photoprotective qH in Arabidopsis
  • 2020
  • In: Nature Plants. - : Nature Publishing Group. - 2055-026X .- 2055-0278. ; 6, s. 154-166
  • Journal article (peer-reviewed)abstract
    • Photosynthetic organisms experience wide fluctuations in light intensity and regulate light harvesting accordingly to prevent damage from excess energy. The antenna quenching component qH is a sustained form of energy dissipation that protects the photosynthetic apparatus under stress conditions. This photoprotective mechanism requires the plastid lipocalin LCNP and is prevented by SUPPRESSOR OF QUENCHING1 (SOQ1) under non-stress conditions. However, the molecular mechanism of qH relaxation has yet to be resolved. Here, we isolated and characterized RELAXATION OF QH1 (ROQH1), an atypical short-chain dehydrogenase–reductase that functions as a qH-relaxation factor in Arabidopsis. The ROQH1 gene belongs to the GreenCut2 inventory specific to photosynthetic organisms, and the ROQH1 protein localizes to the chloroplast stroma lamellae membrane. After a cold and high-light treatment, qH does not relax in roqh1 mutants and qH does not occur in leaves overexpressing ROQH1. When the soq1 and roqh1 mutations are combined, qH can neither be prevented nor relaxed and soq1 roqh1 displays constitutive qH and light-limited growth. We propose that LCNP and ROQH1 perform dosage-dependent, antagonistic functions to protect the photosynthetic apparatus and maintain light-harvesting efficiency in plants.
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9.
  • Barman, Malin, 1983, et al. (author)
  • Short-chain fatty acids (SCFA) in infants’ plasma and corresponding mother's milk and plasma in relation to subsequent sensitisation and atopic disease
  • 2024
  • In: EBioMedicine. - 2352-3964. ; 101
  • Journal article (peer-reviewed)abstract
    • Background: Short-chain fatty acids (SCFAs) in intestinal contents may influence immune function, while less is known about SCFAs in blood plasma. The aims were to investigate the relation between infants’ and maternal plasma SCFAs, as well as SCFAs in mother's milk, and relate SCFA concentrations in infant plasma to subsequent sensitisation and atopic disease. Methods: Infant plasma (N = 148) and corresponding mother's milk and plasma were collected four months postpartum. Nine SCFA (formic, acetic, propionic, isobutyric, butyric, succinic, valeric, isovaleric, and caproic acid) were analysed by UPLC-MS. At 12 months of age, atopic disease was diagnosed by a pediatric allergologist, and sensitisation was measured by skin prick test. All families participated in the Swedish birth cohort NICE (Nutritional impact on Immunological maturation during Childhood in relation to the Environment). Findings: Infants with sensitisation, atopic eczema, or food allergy had significantly lower concentrations of five, three, and two SCFAs, respectively, in plasma at four months. Logistic regressions models showed significant negative associations between formic, succinic, and caproic acid and sensitisation [ORadj (95% CI) per SD: 0.41 (0.19–0.91); 0.19 (0.05–0.75); 0.25 (0.09–0.66)], and between acetic acid and atopic eczema [0.42 (0.18–0.95)], after adjusting for maternal allergy. Infants’ and maternal plasma SCFA concentrations correlated strongly, while milk SCFA concentrations were unrelated to both. Butyric and caproic acid concentrations were enriched around 100-fold, and iso-butyric and valeric acid around 3-5-fold in mother's milk, while other SCFAs were less prevalent in milk than in plasma. Interpretation: Butyric and caproic acid might be actively transported into breast milk to meet the needs of the infant, although mechanistic studies are needed to confirm this. The negative associations between certain SCFAs on sensitisation and atopic disease adds to prior evidence regarding their immunoregulatory potential. Funding: Swedish Research Council (Nr. 2013-3145 and 2019-0137 to A-S.S.), Swedish Research Council for Health, Working Life and Welfare FORTE, Nr 2018-00485 to A.W.), The Swedish Asthma and Allergy Association's Research Fund (2020-0020 to A.S.).
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10.
  • Fristedt, Rikard, et al. (author)
  • A Protein Phosphorylation Threshold for Functional Stacking of Plant Photosynthetic Membranes
  • 2010
  • In: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 5:6
  • Journal article (peer-reviewed)abstract
    • Phosphorylation of photosystem II (PSII) proteins affects macroscopic structure of thylakoid photosynthetic membranes in chloroplasts of the model plant Arabidopsis. In this study, light-scattering spectroscopy revealed that stacking of thylakoids isolated from wild type Arabidopsis and the mutant lacking STN7 protein kinase was highly influenced by cation (Mg++) concentrations. The stacking of thylakoids from the stn8 and stn7stn8 mutants, deficient in STN8 kinase and consequently in light-dependent phosphorylation of PSII, was increased even in the absence of Mg++. Additional PSII protein phosphorylation in wild type plants exposed to high light enhanced Mg++-dependence of thylakoid stacking. Protein phosphorylation in the plant leaves was analyzed during day, night and prolonged darkness using three independent techniques: immunoblotting with anti-phosphothreonine antibodies; Diamond ProQ phosphoprotein staining; and quantitative mass spectrometry of peptides released from the thylakoid membranes by trypsin. All assays revealed dark/night-induced increase in phosphorylation of the 43 kDa chlorophyll-binding protein CP43, which compensated for decrease in phosphorylation of the other PSII proteins in wild type and stn7, but not in the stn8 and stn7stn8 mutants. Quantitative mass spectrometry determined that every PSII in wild type and stn7 contained on average 2.5 +/- 0.1 or 1.4 +/- 0.1 phosphoryl groups during day or night, correspondingly, while less than every second PSII had a phosphoryl group in stn8 and stn7stn8. It is postulated that functional cation-dependent stacking of plant thylakoid membranes requires at least one phosphoryl group per PSII, and increased phosphorylation of PSII in plants exposed to high light enhances stacking dynamics of the photosynthetic membranes.
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11.
  • Fristedt, Rikard, et al. (author)
  • Differential phosphorylation of thylakoid proteins in mesophyll and bundle sheath chloroplasts from maize plants grown under low or high light
  • 2012
  • In: Proteomics. - : Wiley-VCH Verlag Berlin. - 1615-9853 .- 1615-9861. ; 12:18, s. 2852-2861
  • Journal article (peer-reviewed)abstract
    • In C4 plants, such as maize, the photosynthetic apparatus is partitioned over two cell types called mesophyll (M) and bundle sheath (BS), which have different structure and specialization of the photosynthetic thylakoid membranes. We characterized protein phosphorylation in thylakoids of the two cell types from maize grown under either low or high light. Western blotting with phosphothreonine antibodies and ProQ phosphostaining detected light-dependent changes in the protein phosphorylation patterns. LC-MS/MS with alternating CID and electron transfer dissociation sequencing of peptide ions mapped 15 protein phosphorylation sites. Phosphorylated D2, CP29, CP26, Lhcb2 proteins, and ATPsynthase were found only in M membranes. A previously unknown phosphorylation site was mapped in phosphoenolpyruvate carboxykinase from the BS cells. Phosphorylation stoichiometry was calculated from the ratios of normalized ion currents for phosphorylated to nonphosphorylated peptide pairs from the D1, D2, CP43, and PbsH proteins of photosystem II (PSII). Every PSII in M thylakoids contained on average 1.5 +/- 0.1 or 2.3 +/- 0.2 phosphoryl groups in plants grown under either low or high light, while in BS membranes the corresponding numbers were 0.25 +/- 0.1 or 0.7 +/- 0.2, respectively. It is suggested that the phosphorylation level, as well as turnover of PSII depend on the structure of thylakoids.
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12.
  • Fristedt, Rikard, et al. (author)
  • High light induced disassembly of photosystem II supercomplexes in Arabidopsis requires STN7-dependent phosphorylation of CP29
  • 2011
  • In: PLOS ONE. - : PloS. - 1932-6203. ; 6:9
  • Journal article (peer-reviewed)abstract
    • Photosynthetic oxidation of water and production of oxygen by photosystem II (PSII) in thylakoid membranes of plant chloroplasts is highly affected by changes in light intensities. To minimize damage imposed by excessive sunlight and sustain the photosynthetic activity PSII, organized in supercomplexes with its light harvesting antenna, undergoes conformational changes, disassembly and repair via not clearly understood mechanisms. We characterized the phosphoproteome of the thylakoid membranes from Arabidopsis thaliana wild type, stn7, stn8 and stn7stn8 mutant plants exposed to high light. The high light treatment of the wild type and stn8 caused specific increase in phosphorylation of Lhcb4.1 and Lhcb4.2 isoforms of the PSII linker protein CP29 at five different threonine residues. Phosphorylation of CP29 at four of these residues was not found in stn7 and stn7stn8 lacking the STN7 protein kinase. Blue native gel electrophoresis followed by immunological and mass spectrometric analyses of the protein complexes revealed that the high light treatment of the wild type caused migration of CP29 from the PSII supercomplexes to PSII dimers and monomers. A similar high-light-induced disassembly of the PSII supercomplexes occurred in stn8, but not in stn7 and stn7stn8. Transfer of the high-light-treated wild type plants to normal light relocated CP29 back to PSII supercomplexes. We postulate that disassembly of the PSII supercomplexes in plants exposed to high light operate via the STN7-kinase-dependent phosphorylation of the linker protein CP29. Disruption of this adaptive mechanism can explain dramatically retarded growth of the stn7 and stn7stn8 mutants under fluctuating normal/high light conditions.
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13.
  • Fristedt, Rikard, et al. (author)
  • Intrinsically unstructured phosphoprotein TSP9 regulates light harvesting in Arabidopsis thaliana
  • 2009
  • In: Biochemistry. - : American Chemical Society (ACS). - 1520-4995 .- 0006-2960. ; 48:2, s. 499-509
  • Journal article (peer-reviewed)abstract
    • Thylakoid-soluble phosphoprotein of 9 kDa, TSP9, is an intrinsically unstructured plant-specific protein [Song, J., et al. (2006) Biochemistry 45, 15633-15643] with unknown function but established associations with light-harvesting proteins and peripheries of both photosystems [Hansson, M., et al. (2007) J. Biol. Chem. 282, 16214-16222]. To investigate the function of this protein, we used a combination of reverse genetics and biochemical and fluorescence measurement methods in Arabidopsis thaliana. Differential gene expression analysis of plants with a T-DNA insertion in the TSP9 gene using an array of 24000 Arabidopsis genes revealed disappearance of high light-dependent induction of a specific set of mostly signaling and unknown proteins. TSP9-deficient plants had reduced levels of in vivo phosphorylation of light-harvesting complex II polypeptides. Recombinant TSP9 was phosphorylated in light by thylakoid membranes isolated from the wild-type and mutant plants lacking STN8 protein kinase but not by the thylakoids deficient in STN7 kinase, essential for photosynthetic state transitions. TSP9-lacking mutant and RNAi plants with downregulation of TSP9 showed reduced ability to perform state transitions. The nonphotochemical quenching of chlorophyll fluorescence at high light intensities was also less efficient in the mutant compared to wild-type plants. Blue native electrophoresis of thylakoid membrane protein complexes revealed that TSP9 deficiency increased relative stability of photosystem II dimers and supercomplexes. It is concluded that TSP9 regulates plant light harvesting acting as a membrane-binding protein facilitating dissociation of light-harvesting proteins from photosystem II.
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14.
  • Fristedt, Rikard, et al. (author)
  • Phosphorylation of Photosystem II Controls Functional Macroscopic Folding of Photosynthetic Membranes in Arabidopsis
  • 2009
  • In: The Plant Cell. - : Oxford University Press (OUP). - 1040-4651 .- 1532-298X. ; 21:12, s. 3950-3964
  • Journal article (peer-reviewed)abstract
    • Photosynthetic thylakoid membranes in plants contain highly folded membrane layers enriched in photosystem II, which uses light energy to oxidize water and produce oxygen. The sunlight also causes quantitative phosphorylation of major photosystem II proteins. Analysis of the Arabidopsis thaliana stn7xstn8 double mutant deficient in thylakoid protein kinases STN7 and STN8 revealed light-independent phosphorylation of PsbH protein and greatly reduced N-terminal phosphorylation of D2 protein. The stn7xstn8 and stn8 mutants deficient in light-induced phosphorylation of photosystem II had increased thylakoid membrane folding compared with wild-type and stn7 plants. Significant enhancement in the size of stacked thylakoid membranes in stn7xstn8 and stn8 accelerated gravity-driven sedimentation of isolated thylakoids and was observed directly in plant leaves by transmission electron microscopy. Increased membrane folding, caused by the loss of light-induced protein phosphorylation, obstructed lateral migration of the photosystem II reaction center protein D1 and of processing protease FtsH between the stacked and unstacked membrane domains, suppressing turnover of damaged D1 in the leaves exposed to highlight. These findings show that the high level of photosystem II phosphorylation in plants is required for adjustment of macroscopic folding of large photosynthetic membranes modulating lateral mobility of membrane proteins and sustained photosynthetic activity.
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15.
  • Fristedt, Rikard, et al. (author)
  • Photosystem II protein 33, a protein conserved in the plastid lineage, is associated with the chloroplast thylakoid membrane and provides stability to photosystem II supercomplexes in Arabidopsis
  • 2015
  • In: Plant Physiology. - : Oxford University Press (OUP). - 0032-0889 .- 1532-2548. ; 167:2, s. 481-492
  • Journal article (peer-reviewed)abstract
    • Photosystem II (PSII) is a multiprotein complex that catalyzes the light-driven water-splitting reactions of oxygenic photosynthesis. Light absorption by PSII leads to the production of excited states and reactive oxygen species that can cause damage to this complex. Here, we describe Arabidopsis (Arabidopsis thaliana) At1g71500, which encodes a previously uncharacterized protein that is a PSII auxiliary core protein and hence is named PHOTOSYSTEM II PROTEIN33 (PSB33). We present evidence that PSB33 functions in the maintenance of PSII-light-harvesting complex II (LHCII) supercomplex organization. PSB33 encodes a protein with a chloroplast transit peptide and one transmembrane segment. In silico analysis of PSB33 revealed a light-harvesting complex-binding motif within the transmembrane segment and a large surface-exposed head domain. Biochemical analysis of PSII complexes further indicates that PSB33 is an integral membrane protein located in the vicinity of LHCII and the PSII CP43 reaction center protein. Phenotypic characterization of mutants lacking PSB33 revealed reduced amounts of PSII-LHCII supercomplexes, very low state transition, and a lower capacity for nonphotochemical quenching, leading to increased photosensitivity in the mutant plants under light stress. Taken together, these results suggest a role for PSB33 in regulating and optimizing photosynthesis in response to changing light levels.
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16.
  • Fristedt, Rikard (author)
  • Regulatory Functions of Protein Phosphorylation in Plant Photosynthetic Membranes
  • 2010
  • Doctoral thesis (other academic/artistic)abstract
    • Oxygenic photosynthesis is the process in plants, algae and cyanobacteria which converts light energy from the sun into carbohydrates and at the same time produces oxygen from water. Both carbohydrates and oxygen are essential to sustain life on earth. Sunlight is thus a necessity for life, but it can also cause severe problems for photosynthetic organisms, which have evolved several remarkable acclimation systems to cope with light fluctuations in the environment. In higher plants the light driven reactions of photosynthesis proceed in the chloroplast thylakoid membranes highly organized into stacked regions of grana and interconnecting stroma  lamellae. The grana structure is thought to provide functional benefits in the processes of acclimation of the photosynthetic apparatus, particularly in the quality control of photosystem II (PSII) were photodamaged PSII is repaired in a stepwise manner. These processes in the thylakoid membranes were suggested to be regulated by reversible phosphorylation of several proteins in PSII and in its light harvesting antennae complexes (LHCII). Two thylakoid protein kinases, called STN8 and STN7, have been previously identified as responsible for the phosphorylation of PSII and LHCII, respectively. However, molecular mechanisms and the exact functions of these protein phosphorylation events remained largely unknown.In this thesis research I have demonstrated that the PSII protein phosphorylation is needed for the maintenance of the thylakoid structure in Arabidopsis thaliana chloroplasts. A big part of the work on characterization of proteins and their phosphorylation has been done using novel mass spectrometry techniques, and we further developed a label-free method for quantitative studies of protein phosphorylation. The phosphorylation of PSII proteins was found to be diurnal regulated and required for maintenance of the cation-dependent functional stacking of the thylakoid membranes. This phosphorylation was further shown to be important for the regulated turnover of the D1 protein of PSII.Phosphorylation of the plant specific TSP9 protein was found to be dependent on STN7 kinase, and plants deficient in TSP9 showed reduced ability to perform the photosynthetic state transitions and to execute thermal dissipation of excess light energy under high light conditions. I also accomplished characterization of the protein phosphorylation in thylakoids from Arabidopsis plants subjected to high light treatment and discovered STN7-dependent phosphorylation of the antenna protein CP29 required for the adaptive disassembly of PSII supercomplexes in conditions of high light stress.
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17.
  • Gmoser, Rebecca, 1990-, et al. (author)
  • From stale bread and brewers spent grain to a new food source using edible filamentous fungi
  • 2020
  • In: Bioengineered Bugs. - : Informa UK Limited. - 2165-5979 .- 2165-5987. ; 11:1, s. 582-598
  • Journal article (peer-reviewed)abstract
    • By-products from the food sector with a high load of organic matter present both a waste-handling problem related to expenses and to the environment, yet also an opportunity. This study aims to increase the value of stale bread and brewers spent grain (BSG) by re-introducing these residues to the food production chain by converting them to new protein-enriched products using the edible filamentous fungi Neurospora intermedia and Rhizopusoryzae. After 6 days of solid state fermentation (at 35°C, with a95% relative humidity and moisture content of 40% in the substrate) on stale bread, a nutrient-rich fungal-fermented product was produced. The total protein content, as analyzed by total amino acids, increased from 16.5% in stale sourdough bread to 21.1% (on dry weight basis) in the final product with an improved relative ratio of essential amino acids. An increase in dietary fiber, minerals (Cu, Fe, Zn) and vitamin E, as well as an addition of vitamin D2 (0.89 µg/g dry weight sample) was obtained compared with untreated stale bread. Furthermore, addition of BSG to the sourdough bread with the aim to improve textural changes after fermentation showed promising outcomes. Cultivation of N. intermedia or R. oryzae on stale sourdough bread mixed with 6.5% or 11.8% BSG, respectively, resulted in fungal-fermented products with similar textural properties to a commercial soybean burger. Bioconversion of stale bread and BSG by fungal solid state fermentation to produce a nutrient-enriched food product was confirmed to be a successful way to minimize food waste and protein shortage.
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18.
  • Ingelsson, Björn, 1983-, et al. (author)
  • Phosphorylation stoichiometry determination in plant photosynthetic membranes.
  • 2015
  • In: Plant Phosphoproteomics. - New York : Springer-Verlag New York. - 9781493926473 - 9781493926480 ; , s. 121-134
  • Book chapter (peer-reviewed)abstract
    • This chapter describes different strategies for the study of phosphorylation dynamics and stoichiometry in photosynthetic membranes. Detailed procedures for the detection, large-scale identification, and quantification of phosphorylated proteins optimized for plant thylakoid proteins are given.
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19.
  • Moparthi, Satish Babu, et al. (author)
  • Chaperone activity of Cyp18 through hydrophobic condensation that enables rescue of transient misfolded molten globule intermediates
  • 2010
  • In: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 49:6, s. 1137-1145
  • Journal article (peer-reviewed)abstract
    • The single-domain cyclophilin 18 (Cyp18) has long been known to function as a peptidyl-prolyl cis/trans isomerase (PPI) and was proposed by us to also function as a chaperone [Freskgård, P.-O., Bergenhem, N., Jonsson, B.-H., Svensson, M., and Carlsson, U. (1992) Science 258, 466−468]. Later several multidomain PPIs were demonstrated to work as both a peptidyl-prolyl cis/trans isomerase and a chaperone. However, the chaperone ability of Cyp18 has been debated. In this work, we add additional results that show that Cyp18 can both accelerate the rate of refolding and increase the yield of native protein during the folding reaction, i.e., function as both a folding catalyst and a chaperone. Refolding experiments were performed using severely destabilized mutants of human carbonic anhydrase II under conditions where the unfolding reaction is significant and a larger fraction of a more destabilized variant populates molten globule-like intermediates during refolding. A correlation of native state protein stability of the substrate protein versus Cyp18 chaperone activity was demonstrated. The induced correction of misfolded conformations by Cyp18 likely functions through rescue from misfolding of transient molten globule intermediates. ANS binding data suggest that the interaction by Cyp18 leads to an early stage condensation of accessible hydrophobic portions of the misfolding-prone protein substrate during folding. The opposite effect was observed for GroEL known as an unfoldase at early stages of refolding. The chaperone effect of Cyp18 was also demonstrated for citrate synthase, suggesting a general chaperone effect of this PPI.
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20.
  • Nilsson, Anders K., 1982, et al. (author)
  • PSB33 protein sustains photosystem II in plant chloroplasts under UV-A light
  • 2020
  • In: Journal of Experimental Botany. - OXFORD ENGLAND : Oxford University Press (OUP). - 0022-0957 .- 1460-2431. ; 71:22, s. 7210-7223
  • Journal article (peer-reviewed)abstract
    • Plants can quickly and dynamically respond to spectral and intensity variations of the incident light. These responses include activation of developmental processes, morphological changes, and photosynthetic acclimation that ensure optimal energy conversion and minimal photoinhibition. Plant adaptation and acclimation to environmental changes have been extensively studied, but many details surrounding these processes remain elusive. The photosystem II (PSII)-associated protein PSB33 plays a fundamental role in sustaining PSII as well as in the regulation of the light antenna in fluctuating light. We investigated how PSB33 knock-out Arabidopsis plants perform under different light qualities. psb33 plants displayed a reduction of 88% of total fresh weight compared to wild type plants when cultivated at the boundary of UV-A and blue light. The sensitivity towards UV-A light was associated with a lower abundance of PSII proteins, which reduces psb33 plants' capacity for photosynthesis. The UV-A phenotype was found to be linked to altered phytohormone status and changed thylakoid ultrastructure. Our results collectively show that PSB33 is involved in a UV-A light-mediated mechanism to maintain a functional PSII pool in the chloroplast.
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21.
  • Oliveira, Gabriel, et al. (author)
  • Extracts of Digested Berries Increase the Survival of Saccharomyces cerevisiae during H2O2 Induced Oxidative Stress
  • 2021
  • In: Molecules. - : MDPI AG. - 1420-3049 .- 1420-3049. ; 26:4
  • Journal article (peer-reviewed)abstract
    • Many studies suggest anthocyanins may prevent the development of several diseases. However, anthocyanin bioactivity against cellular stress is not fully understood. This study aimed to evaluate the protective effect of berry anthocyanins on stressed cells using Saccharomyces cerevisiae. The impact of in vitro gastrointestinal digestion on anthocyanin profiles was also assessed. Bilberry and blackcurrant had higher anthocyanin levels than raspberry and strawberry, but digestion reduced the detected anthocyanins by approximately 90%. Yeast cells with and without digested or nondigested anthocyanin extracts were exposed to H2O2 and examined for survival. In the presence of anthocyanins, particularly from digested strawberry, a significant increase in cell survival was observed, suggesting that the type and levels of anthocyanins are important factors, but they also need to undergo gastrointestinal (GI) structural modifications to induce cell defence. Results also showed that cells need to be exposed to anthocyanins before the stress was applied, suggesting induction of a cellular defence system by anthocyanins or their derivatives rather than by a direct antioxidative effect on H2O2. Overall, data showed that exposure of severely stressed yeast cells to digested berry extracts improved cell survival. The findings also showed the importance of considering gastrointestinal digestion when evaluating anthocyanins' biological activity.
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22.
  • Romanowska, Elzbieta, et al. (author)
  • Phosphorylation of PSII proteins in maize thylakoids in the presence of Pb ions
  • 2012
  • In: Journal of plant physiology (Print). - : Elsevier. - 0176-1617 .- 1618-1328. ; 169:4, s. 345-352
  • Journal article (peer-reviewed)abstract
    • Lead is potentially toxic to all organisms including plants. Many physiological studies suggest that plants have developed various mechanisms to contend with heavy metals, however the molecular mechanisms remain unclear. We studied maize plants in which lead was introduced into detached leaves through the transpiration stream. The photochemical efficiency of PSII, measured as an Fv/Fm ratio, in the maize leaves treated with Pb was only 10% lower than in control leaves. The PSII activity was not affected by Pb ions in mesophyll thylakoids, whereas in bundle sheath it was reduced. Protein phosphorylation in mesophyll and bundle sheath thylakoids was analyzed using mass spectrometry and protein blotting before and after lead treatment. Both methods clearly demonstrated increase in phosphorylation of the PSII proteins upon treatment with Pb2+, however, the extent of D1, D2 and CP43 phosphorylation in the mesophyll chloroplasts was clearly higher than in bundle sheath cells. We found that in the presence of Pb ions there was no detectable dephosphorylation of the strongly phosphorylated D1 and PsbH proteins of PSII complex in darkness or under far red light. These results suggest that Pb2+ stimulates phosphorylation of PSII core proteins, which can affect stability of the PSII complexes and the rate of D1 protein degradation. Increased phosphorylation of the PSII core proteins induced by Pb ions may be a crucial protection mechanism stabilizing optimal composition of the PSII complexes under metal stress conditions. Our results show that acclimation to Pb ions was achieved in both types of maize chloroplasts in the same way. However, these processes are obviously more complex because of different metabolic status in mesophyll and bundle sheath chloroplasts.
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23.
  • Rousta, Neda, et al. (author)
  • Production of fungal biomass from oat flour for the use as a nutritious food source
  • 2022
  • In: NFS Journal. - : Elsevier BV. - 2352-3646. ; 29, s. 8-15
  • Journal article (peer-reviewed)abstract
    • Fermentation can be a powerful tool for developing new sustainable foods with increased nutritional value and fermented microbial biomass derived from filamentous fungi is a promising example. This study investigates the nutritional profile of edible Aspergillus oryzae biomass produced under submerged fermentation (SmF) using oat flour as a substrate. The fermentation occurred in a 1m3 airlift bioreactor during 48 h at 35 °C and the nutritional profile of the produced fungal biomass in terms of amino acids, fatty acids, minerals (Fe, Zn, Cu, Mn), vitamins (E, D2), and dietary fiber was compared to oat flour as well as pure fungal biomass grown on semi-synthetic medium. The total amount of amino acids increased from 11% per dry weight (dw) in oat flour to 23.5% dw in oat fungal biomass with an improved relative ratio of essential amino acids (0.37 to 0.42). An increase in dietary fibers, minerals (Fe, Zn, Cu), vitamin E, as well as vitamin D2 were also obtained in the oat fungal biomass compared to oat flour. Moreover, the short chain omega-3 α-linolenic acid (ALA) and omega-6 linoleic acid (LA) values increased from 0.6 to 8.4 and 21.7 to 68.4 (mg/g dry weight sample), respectively, in oat fungal biomass. The results indicate that fungal biomass grown on oat flour could have a potential application in the food industry as a nutritious source for a wide variety of products. 
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24.
  • Sajib, Mursalin, 1987, et al. (author)
  • Hemoglobin-mediated lipid oxidation of herring filleting co-products during ensilaging and its inhibition by pre-incubation in antioxidant solutions
  • 2021
  • In: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322 .- 2045-2322. ; 11:1, s. 1-12
  • Journal article (peer-reviewed)abstract
    • The aims of this study were to investigate the role of hemoglobin (Hb) in lipid oxidation development during ensilaging of herring filleting co-products, and, to inhibit this reaction by pre-incubating the co-products in water or physiological salt, with/without different antioxidants. Results showed that both peroxide value (PV) and 2-thiobarbituric acid reactive substances (TBARS) gradually increased during 7 days of ensilaging at 22 °C in absence of antioxidants. The increase in TBARS was proportional to the Hb levels present, while PV was less affected. A Hb-fortified Tris-buffer model system adjusted to pH 3.50 confirmed that Hb changed immediately from its native oxyHb to the metHb state, which facilitated heme group release and thus probably explains the increased PV and TBARS during ensilaging. Pre-incubating the co-products for 30 s in a solution containing 0.5% rosemary extract was the most promising strategy to inhibit lipid oxidation both in the co-products during pre-processing storage and during the actual ensilaging. The solution could be re-used up to ten times without losing its activity, illustrating that this methodology can be a scalable and cost-effective strategy to extend the oxidative stability of herring co-products allowing for further value adding e.g., into a high-quality silage.
  •  
25.
  • Sar, Taner, Postdoctoral Researcher, 1989-, et al. (author)
  • Demo-scale production of protein-rich fungal biomass from potato protein liquor for use as innovative food and feed products
  • 2022
  • In: Food Bioscience. - : Elsevier BV. - 2212-4292 .- 2212-4306. ; 47
  • Journal article (peer-reviewed)abstract
    • Innovative food and feed products have recently attracted the attention of both producers and consumers. Filamentous fungi are important biomass producers with their high protein contents. In this study, fungal biomass production from edible potato protein liquor (PPL), generated during starch production processes, was investigated through different fungal strains (Rhizopus oryzae, R. oligosporus, R. delemar, Aspergillus oryzae and Neurospora intermedia). The effects of PPL concentration, incubation time, initial pH, and cultivation conditions (in shake flaks and different scale reactors) were examined to determine the amount of biomass and its crude protein level. It was determined that the fungal biomass produced by R. delemar in industrial scale contained 53% crude protein. For this strain, the amino acid and fatty acid profiles as well as metals (iron, manganese, copper, and zinc) of the produced biomass were also investigated to assess possible use as a food or feed source. The R. delemar fungal biomass can be a promising raw material for feed and food production, for example, considering its protein and fatty acid profiles with 41% essential amino acids and 33% polyunsaturated fatty acids.
  •  
26.
  • Sirpio, Sari, et al. (author)
  • AtCYP38 ensures early biogenesis, correct assembly and sustenance of photosystem II
  • 2008
  • In: The Plant Journal. - 0960-7412 .- 1365-313X. ; 55:4, s. 639-651
  • Journal article (peer-reviewed)abstract
    • AtCYP38 is a thylakoid lumen protein comprising the immunophilin domain and the phosphatase inhibitor module. Here we show the association of AtCYP38 with the photosystem II (PSII) monomer complex and address its functional role using AtCYP38-deficient mutants. The dynamic greening process of etiolated leaves failed in the absence of AtCYP38, due to specific problems in the biogenesis of PSII complexes. Also the development of leaves under short-day conditions was severely disturbed. Detailed biophysical and biochemical analysis of mature AtCYP38-deficient plants from favorable growth conditions (long photoperiod) revealed: (i) intrinsic malfunction of PSII, which (ii) occurred on the donor side of PSII and (iii) was dependent on growing light intensity. AtCYP38 mutant plants also showed decreased accumulation of PSII, which was shown not to originate from impaired D1 synthesis or assembly of PSII monomers, dimers and supercomplexes as such but rather from the incorrect fine-tuning of the oxygen-evolving side of PSII. This, in turn, rendered PSII centers extremely susceptible to photoinhibition. AtCYP38 deficiency also drastically decreased the in vivo phosphorylation of PSII core proteins, probably related to the absence of the AtCYP38 phosphatase inhibitor domain. It is proposed that during PSII assembly AtCYP38 protein guides the proper folding of D1 (and CP43) into PSII, thereby enabling the correct assembly of the water-splitting Mn 4-Ca cluster even with high turnover of PSII. © 2008 The Authors.
  •  
27.
  • Wang, Ricky, et al. (author)
  • In vitro protein digestibility of edible filamentous fungi compared to common food protein sources
  • 2023
  • In: Food Bioscience. - : Elsevier. - 2212-4306 .- 2212-4292. ; 54
  • Journal article (peer-reviewed)abstract
    • Edible filamentous fungi, as a source of mycoprotein, is an emerging sustainable protein source as it can be cultivated on food-industry sidestreams, thus providing the food system with circularity. However, the digestibility of mycoprotein from different species of fungi is yet to be studied and compared to commonly consumed food proteins derived from muscle. Using the static INFOGEST in vitro gastrointestinal (GI) digestion protocol, but with less pancreatin than the recommended amount to omit high background from enzyme autolysis, this study investigated the protein degree of hydrolysis (DH%) and amino acid accessibility of five species of edible fungi in comparison with salmon fillet, chicken breast, beef tenderloin and casein. Three of the edible fungi species reached protein DH% between 58% ± 2.6% and 62% ± 5.6% during GI digestion compared to chicken, salmon, and beef reaching 62%–67% as well as casein at 55%. The amino acid accessibility of fungi (81%–92%), was comparable to that of salmon, chicken breast, and beef (90%–94%). This study thus indicated that edible fungi is a sustainable and nutritionally sound protein source.
  •  
28.
  • Wu, Haizhou, 1987, et al. (author)
  • Pilot-Scale Antioxidant Dipping of Herring (Clupea harengus) Co-products to Allow Their Upgrading to a High-Quality Mince for Food Production
  • 2023
  • In: ACS Sustainable Chemistry & Engineering. - : American Chemical Society (ACS). - 2168-0485. ; 11:12, s. 4727-4737
  • Journal article (peer-reviewed)abstract
    • To enable production of high-quality mince from herring backbones, a scalable antioxidant strategy is needed due to the high susceptibility of herring muscle to lipid oxidation. We here measured the stabilizing effect of lab-/pilot-scale predipping of herring backbones (30-500 kg) in antioxidant solutions prior to production of mechanically separated mince (MSM). The antioxidants were (i) Duralox MANC, a mixture of rosemary extract, ascorbic acid, α-tocopherol, and citric acid, and (ii) rosemary extract with or without isoascorbic acid. Delivery of the key rosemary-derived antioxidant components carnosol and carnosic acid was monitored during the dipping process and ice/frozen storage. Predipping in 2% Duralox MANC gave MSM with 26.7-31.7 mg/kg carnosol + carnosic acid and extended the oxidation lag phase from <1 to 12 days during ice storage and from <1 to 6 months during frozen storage compared to control. Dipping in 0.2% rosemary extract with or without 0.5% isoascorbic acid solution gave MSM with 20.6-28.2 mg/kg carnosol + carnosic acid and extended the lag phase to 6 days and 9 months during ice and frozen storage, respectively. Our results confirmed, in pilot scale, that predipping herring coproducts in antioxidant solutions is a promising strategy to utilize these raw materials for, e.g., mince and burger production rather than for low value products as fish meal.
  •  
29.
  • Yin, Lan, 1979, et al. (author)
  • Photosystem II Function and Dynamics in Three Widely Used Arabidopsis thaliana Accessions
  • 2012
  • In: PLoS One. - : Public Library of Science (PLoS). - 1932-6203. ; 7:9
  • Journal article (peer-reviewed)abstract
    • Columbia-0 (Col-0), Wassilewskija-4 (Ws-4), and Landsberg erecta-0 (Ler-0) are used as background lines for many public Arabidopsis mutant collections, and for investigation in laboratory conditions of plant processes, including photosynthesis and response to high-intensity light (HL). The photosystem II (PSII) complex is sensitive to HL and requires repair to sustain its function. PSII repair is a multistep process controlled by numerous factors, including protein phosphorylation and thylakoid membrane stacking. Here we have characterized the function and dynamics of PSII complex under growth-light and HL conditions. Ws-4 displayed 30% more thylakoid lipids per chlorophyll and 40% less chlorophyll per carotenoid than Col-0 and Ler-0. There were no large differences in thylakoid stacking, photoprotection and relative levels of photosynthetic complexes among the three accessions. An increased efficiency of PSII closure was found in Ws-4 following illumination with saturation flashes or continuous light. Phosphorylation of the PSII D1/D2 proteins was reduced by 50% in Ws-4 as compared to Col-0 and Ler-0. An increase in abundance of the responsible STN8 kinase in response to HL treatment was found in all three accessions, but Ws-4 displayed 50% lower levels than Col-0 and Ler-0. Despite this, the HL treatment caused in Ws-4 the lagest extent of PSII inactivation, disassembly, D1 protein degradation, and the largest decrease in the size of stacked thylakoids. The dilution of chlorophyll-protein complexes with additional lipids and carotenoids in Ws-4 may represent a mechanism to facilitate lateral protein traffic in the membrane, thus compensating for the lack of a full complement of STN8 kinase. Nevertheless, additional PSII damage occurs in Ws-4, which exceeds the D1 protein synthesis capacity, thus leading to enhanced photoinhibition. Our findings are valuable for selection of appropriate background line for PSII characterization in Arabidopsis mutants, and also provide the first insights into natural variation of PSII protein phosphorylation.
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