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T N Popova - One of the best experts on this subject based on the ideXlab platform.
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influence of 10 6 plastoquinonyl decyltriphenylphosphonium skq1 on oxidative status in rats with protamine sulfate induced hyperglycemia
Biochemistry, 2015Co-Authors: Ya G Voronkova, T N Popova, A A Agarkov, M V SkulachevAbstract:An influence of 10-(6'-plastoquinonyl)decyltriphenylphosphonium (SkQ1) on oxidative status and activity of some antioxidant enzymes in the liver and blood serum from rats was examined during experimental hyperglycemia developed after injecting protamine sulfate. It was found that SkQ1 lowered glycemic level in rats treated with protamine sulfate. Moreover, it was also accompanied by restoration of the normal range of biochemiluminescence parameters indicating the rate of ongoing free radical processes, magnitude of primary products of lipid peroxidation such as diene conjugates, activity of Aconitate Hydratase, and level of citrate in rat liver and blood. Hence, it was demonstrated that activity of superoxide dismutase and catalase, increasing during hyperglycemia, was decreased after administering SkQ1. This might be related to the ability of SkQ1 to normalize free-radical homeostasis imbalanced during hyperglycemia.
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Effect of Succinic Acid and Chitosan Derivatives on Aconitate Hydratase Activity and Citrate Content in Rat Brain Tissues Under Ischemia/Reperfusion Conditions
Pharmaceutical Chemistry Journal, 2015Co-Authors: O. A. Safonova, T N Popova, A. I. Slivkin, A. S. Belenova, Yu. TalmiAbstract:Succinic-acid and chitosan derivatives administered at various dose to rats with brain ischemia/reperfusion led to a decrease in the level of lactate (an ischemia marker) in brain tissue as compared to the value in pathology. The activity of Aconitate Hydratase (a sensitive target of free-radical action) increased under these conditions whereas the citrate level in the brain and blood serum of the animals decreased toward the control values. The observed changes were dose-dependent. The results showed that these drugs were capable of reducing the degree of metabolic damage and the development of oxidative stress during post-ischemic reperfusion. Therefore, the investigated substances may be of considerable interest for pharmacological correction of metabolic changes during the development of such pathologies. Chitosan succinate exhibited more pronounced neuroprotective and antioxidant effects than N -succinylchitosan.
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molecular biological problems of drug design and mechanism of drug action effect of succinic acid and chitosan derivatives on Aconitate Hydratase activity and citrate content in rat brain tissues under ischemia reperfusion conditions
2015Co-Authors: O. A. Safonova, T N Popova, A. I. Slivkin, A. S. BelenovaAbstract:Succinic-acid and chitosan derivatives administered at various dose to rats with brain ischemia/reperfusion led to a decrease in the level of lactate (an ischemia marker) in brain tissue as compared to the value in pathology. The activity of Aconitate Hydratase (a sensitive target of free-radical action) increased under these conditions whereas the citrate level in the brain and blood serum of the animals decreased toward the control values. The observed changes were dose-dependent. The results showed that these drugs were capable of reducing the degree of metabolic damage and the development of oxidative stress during post-ischemic reperfusion. Therefore, the investigated substances may be of considerable interest for pharmacological correction of metabolic changes during the development of such pathologies. Chitosan succinate exhibited more pronounced neuroprotective and antioxidant effects than N-succinylchitosan.
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effect of melaxen and valdoxan on free radical processes intensity Aconitate Hydratase activity and citrate content in rats tissues under hyperthyroidism
Biomeditsinskaya khimiya, 2014Co-Authors: M V Gorbenko, T N Popova, K K Shulgin, S S Popov, A A AgarkovAbstract:The influence of melaxen and valdoxan on the biochemiluminescence parameters, Aconitate Hydratase activity and citrate level in rats heart and liver during development of experimental hyperthyroidism has been investigated. Administration of these substances promoted a decrease of biochemiluminescence parameters, which had been increased in tissues of rats in response to the development of oxidative stress under hyperthyroidism. Aconitate Hydratase activity and citrate concentration in rats liver and heart, growing at pathological conditions, changed towards control value after administration of the drugs correcting melatonin level. The results indicate the positive effect of valdoxan and melaxen on oxidative status of the organism under the development of experimental hyperthyroidism that is associated with antioxidant action of melatonin.
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Aconitate Hydratase of mammals under oxidative stress
Biochemistry (Moscow), 2008Co-Authors: L V Matasova, T N PopovaAbstract:Data on the structure, functions, regulation of activity, and expression of cytosolic and mitochondrial Aconitate Hydratase isoenzymes of mammals are reviewed. The role of Aconitate Hydratase and structurally similar iron-regulatory protein in maintenance of homeostasis of cell iron is described. Information on modifications of the Aconitate Hydratase molecule and changes in expression under oxidative stress is generalized. The role of Aconitate Hydratase in the pathogenesis of some diseases is considered.
Anchalee Tungtrongchitr - One of the best experts on this subject based on the ideXlab platform.
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proteome allergenome and novel allergens of house dust mite dermatophagoides farinae
Journal of Proteome Research, 2016Co-Authors: Jintarat Choopong, Onrapak Reamtong, Nitat Sookrung, Watee Seesuay, Nitaya Indrawattana, Yuwaporn Sakolvaree, Wanpen Chaicumpa, Anchalee TungtrongchitrAbstract:Dermatophagoides farinae mite is a predominant source of indoor allergens causing high incidence of allergy worldwide. People with different genetic background respond differently to the mite components, and thus the component-resolved diagnosis (CRD) is preferred to the conventional allergy test based on crude mite extract. In this study, proteome and culprit components in the D. farinae whole body extract that sensitized the allergic patients were studied by using SDS-PAGE (1DE) and 2DE-IgE immunoblotting followed by LC-MS/MS and database search for protein identification. From the 1DE, the mite extract revealed 105 proteins that could be classified into seven functionally different groups: allergens, structural components, enzymes, enzyme inhibitor, receptor proteins, transporters, and binding/regulatory/cell signaling proteins. From the 2DE, the mite extract produced 94 spots; 63 were bound by IgE in sera of 20 D. farinae allergic patients. One more protein that was not revealed by the 2DE and protein staining reacted with IgE in 2 allergic patients. Proteins in 40 spots could be identified as 35 different types. Three of them reacted to IgE of >50% of the allergic patients, and hence they are major allergens: tropomyosin or Der f 10 (75%), Aconitate Hydratase (70%), and one uncharacterized protein (55%). Aconitate Hydratase is a novel D. farinae major allergen unraveled in this study. Several mite minor allergens that have never been previously reported are also identified. The data have clinical applications in the component-resolved diagnosis for tailor-designed allergen-specific immunotherapy.
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Proteome, Allergenome, and Novel Allergens of House Dust Mite, Dermatophagoides farinae
2016Co-Authors: Jintarat Choopong, Onrapak Reamtong, Nitat Sookrung, Watee Seesuay, Nitaya Indrawattana, Yuwaporn Sakolvaree, Wanpen Chaicumpa, Anchalee TungtrongchitrAbstract:Dermatophagoides farinae mite is a predominant source of indoor allergens causing high incidence of allergy worldwide. People with different genetic background respond differently to the mite components, and thus the component-resolved diagnosis (CRD) is preferred to the conventional allergy test based on crude mite extract. In this study, proteome and culprit components in the D. farinae whole body extract that sensitized the allergic patients were studied by using SDS-PAGE (1DE) and 2DE-IgE immunoblotting followed by LC–MS/MS and database search for protein identification. From the 1DE, the mite extract revealed 105 proteins that could be classified into seven functionally different groups: allergens, structural components, enzymes, enzyme inhibitor, receptor proteins, transporters, and binding/regulatory/cell signaling proteins. From the 2DE, the mite extract produced 94 spots; 63 were bound by IgE in sera of 20 D. farinae allergic patients. One more protein that was not revealed by the 2DE and protein staining reacted with IgE in 2 allergic patients. Proteins in 40 spots could be identified as 35 different types. Three of them reacted to IgE of >50% of the allergic patients, and hence they are major allergens: tropomyosin or Der f 10 (75%), Aconitate Hydratase (70%), and one uncharacterized protein (55%). Aconitate Hydratase is a novel D. farinae major allergen unraveled in this study. Several mite minor allergens that have never been previously reported are also identified. The data have clinical applications in the component-resolved diagnosis for tailor-designed allergen-specific immunotherapy
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Proteome and Allergenome of Asian Wasp, Vespa affinis, Venom and IgE Reactivity of the Venom Components
2014Co-Authors: Nitat Sookrung, Onrapak Reamtong, Nitaya Indrawattana, Yuwaporn Sakolvaree, Anchalee Tungtrongchitr, Siriporn Wong-din-dam, Nualanong Visitsunthorn, Wiparat Manuyakorn, Wanpen ChaicumpaAbstract:Vespa affinis (Asian wasp, Thai banded tiger wasp, or local name: Tor Hua Seua) causes the most frequent incidence of medically important Hymenoptera sting in South and Southeast Asia. However, data on the venom components attributable to the sting derived-clinical manifestations (local reactions, IgE mediated-anaphylaxis, or systemic envenomation) are lacking. This study provides the first set information on V. affinis venom proteome, allergenome, and IgE reactivity of individual venom components. From 2DE-gel based-proteomics, the venom revealed 93 protein spots, of which proteins in 51 spots could be identified and classified into three groups: typical venom components and structural and housekeeping proteins. Venom proteins in 32 spots reacted with serum IgE of wasp allergic patients. Major allergenic proteins that reacted to IgE of >50% of the wasp allergic patients included PLA1 (100%), arginine kinase (73%), heat shock 70 kDa protein (73.3%), venom allergen-5 (66.7%), enolase (66.7%), PLA1 magnifin (60%), glyceraldehyde-3-phosphate dehydrogenase (60%), hyaluronidase (53.3%), and fructose-bisphosphate aldolase (53.3%). The venom minor allergens were GB17876 transcript (40%), GB17291 transcript (20%), malic enzyme (13.3%), Aconitate Hydratase (6.7%), and phosphoglucomutase (6.7%). The information has diagnostic and clinical implications for future improvement of case diagnostic sensitivity and specificity, component-resolve diagnosis, and design of specific Hymenoptera venom immunotherapy
Onrapak Reamtong - One of the best experts on this subject based on the ideXlab platform.
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Adaptive effect of sericin on hepatic mitochondrial conformation through its regulation of apoptosis, autophagy and energy maintenance: a proteomics approach.
Scientific Reports, 2018Co-Authors: Sumate Ampawong, Duangnate Isarangkul, Onrapak Reamtong, Pornanong AramwitAbstract:We recently demonstrated that in addition to its protective effect on pancreatic and adrenal biosynthesis, antioxidant properties of sericin decrease blood cholesterol levels and improve the liver mitochondrial architecture. However, little is known about the detailed mechanisms underlying these effects. Using proteomics and electron microscopy, we identified mitochondrial proteins that play important roles in the preservation of the mitochondrial ultrastructure and cholesterol-lowering properties of sericin. Our results showed that sericin maintains the mitochondrial architecture during conditions of high blood cholesterol by regulating apoptotic (NADH-ubiquinone oxidoreductase 75 kDa subunit) and autophagic (mitochondrial elongation factor Tu and prohibitin-2) proteins as well as energy maintenance proteins [haloacid dehalogenase-like hydrolase domain-containing protein 3, succinate dehydrogenase (ubiquinone) flavoprotein subunit, ATP synthase-α subunit precursor, enoyl-CoA Hydratase domain-containing protein 3 and electron transfer flavoprotein subunit-α]. Sericin also exerts anti-oxidative properties via Aconitate Hydratase and Chain A, crystal structure of rat carnitine palmitoyltrasferase 2 proteins. Together, these activities may reduce hepatocytic triglyceride deposition, thereby decreasing steatosis, as demonstrated by the modulatory effects on ornithine aminotransferase, mitochondrial aspartate aminotransferase, acyl-CoA synthase, hydroxyacyl-CoA dehydrogenase and D-beta-hydroxybutyrate dehydrogenase. Sericin activity further balanced nitrogenous waste detoxification, characterised by carbamoyl-phosphate synthase (ammonia), aldehyde dehydrogenase and uricase, or folate biosynthesis via sarcosine dehydrogenase and dimethyl glycine dehydrogenase. These results suggest that sericin maintains the hepatic mitochondrial architecture through apoptotic, autophagic, energy maintenance and anti-oxidative mitochondrial proteins for alleviating hepatic steatosis and promoting liver function under conditions of hypercholesterolaemia.
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proteome allergenome and novel allergens of house dust mite dermatophagoides farinae
Journal of Proteome Research, 2016Co-Authors: Jintarat Choopong, Onrapak Reamtong, Nitat Sookrung, Watee Seesuay, Nitaya Indrawattana, Yuwaporn Sakolvaree, Wanpen Chaicumpa, Anchalee TungtrongchitrAbstract:Dermatophagoides farinae mite is a predominant source of indoor allergens causing high incidence of allergy worldwide. People with different genetic background respond differently to the mite components, and thus the component-resolved diagnosis (CRD) is preferred to the conventional allergy test based on crude mite extract. In this study, proteome and culprit components in the D. farinae whole body extract that sensitized the allergic patients were studied by using SDS-PAGE (1DE) and 2DE-IgE immunoblotting followed by LC-MS/MS and database search for protein identification. From the 1DE, the mite extract revealed 105 proteins that could be classified into seven functionally different groups: allergens, structural components, enzymes, enzyme inhibitor, receptor proteins, transporters, and binding/regulatory/cell signaling proteins. From the 2DE, the mite extract produced 94 spots; 63 were bound by IgE in sera of 20 D. farinae allergic patients. One more protein that was not revealed by the 2DE and protein staining reacted with IgE in 2 allergic patients. Proteins in 40 spots could be identified as 35 different types. Three of them reacted to IgE of >50% of the allergic patients, and hence they are major allergens: tropomyosin or Der f 10 (75%), Aconitate Hydratase (70%), and one uncharacterized protein (55%). Aconitate Hydratase is a novel D. farinae major allergen unraveled in this study. Several mite minor allergens that have never been previously reported are also identified. The data have clinical applications in the component-resolved diagnosis for tailor-designed allergen-specific immunotherapy.
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Proteome, Allergenome, and Novel Allergens of House Dust Mite, Dermatophagoides farinae
2016Co-Authors: Jintarat Choopong, Onrapak Reamtong, Nitat Sookrung, Watee Seesuay, Nitaya Indrawattana, Yuwaporn Sakolvaree, Wanpen Chaicumpa, Anchalee TungtrongchitrAbstract:Dermatophagoides farinae mite is a predominant source of indoor allergens causing high incidence of allergy worldwide. People with different genetic background respond differently to the mite components, and thus the component-resolved diagnosis (CRD) is preferred to the conventional allergy test based on crude mite extract. In this study, proteome and culprit components in the D. farinae whole body extract that sensitized the allergic patients were studied by using SDS-PAGE (1DE) and 2DE-IgE immunoblotting followed by LC–MS/MS and database search for protein identification. From the 1DE, the mite extract revealed 105 proteins that could be classified into seven functionally different groups: allergens, structural components, enzymes, enzyme inhibitor, receptor proteins, transporters, and binding/regulatory/cell signaling proteins. From the 2DE, the mite extract produced 94 spots; 63 were bound by IgE in sera of 20 D. farinae allergic patients. One more protein that was not revealed by the 2DE and protein staining reacted with IgE in 2 allergic patients. Proteins in 40 spots could be identified as 35 different types. Three of them reacted to IgE of >50% of the allergic patients, and hence they are major allergens: tropomyosin or Der f 10 (75%), Aconitate Hydratase (70%), and one uncharacterized protein (55%). Aconitate Hydratase is a novel D. farinae major allergen unraveled in this study. Several mite minor allergens that have never been previously reported are also identified. The data have clinical applications in the component-resolved diagnosis for tailor-designed allergen-specific immunotherapy
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Proteome and Allergenome of Asian Wasp, Vespa affinis, Venom and IgE Reactivity of the Venom Components
2014Co-Authors: Nitat Sookrung, Onrapak Reamtong, Nitaya Indrawattana, Yuwaporn Sakolvaree, Anchalee Tungtrongchitr, Siriporn Wong-din-dam, Nualanong Visitsunthorn, Wiparat Manuyakorn, Wanpen ChaicumpaAbstract:Vespa affinis (Asian wasp, Thai banded tiger wasp, or local name: Tor Hua Seua) causes the most frequent incidence of medically important Hymenoptera sting in South and Southeast Asia. However, data on the venom components attributable to the sting derived-clinical manifestations (local reactions, IgE mediated-anaphylaxis, or systemic envenomation) are lacking. This study provides the first set information on V. affinis venom proteome, allergenome, and IgE reactivity of individual venom components. From 2DE-gel based-proteomics, the venom revealed 93 protein spots, of which proteins in 51 spots could be identified and classified into three groups: typical venom components and structural and housekeeping proteins. Venom proteins in 32 spots reacted with serum IgE of wasp allergic patients. Major allergenic proteins that reacted to IgE of >50% of the wasp allergic patients included PLA1 (100%), arginine kinase (73%), heat shock 70 kDa protein (73.3%), venom allergen-5 (66.7%), enolase (66.7%), PLA1 magnifin (60%), glyceraldehyde-3-phosphate dehydrogenase (60%), hyaluronidase (53.3%), and fructose-bisphosphate aldolase (53.3%). The venom minor allergens were GB17876 transcript (40%), GB17291 transcript (20%), malic enzyme (13.3%), Aconitate Hydratase (6.7%), and phosphoglucomutase (6.7%). The information has diagnostic and clinical implications for future improvement of case diagnostic sensitivity and specificity, component-resolve diagnosis, and design of specific Hymenoptera venom immunotherapy
L V Matasova - One of the best experts on this subject based on the ideXlab platform.
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Aconitate Hydratase of mammals under oxidative stress
Biochemistry (Moscow), 2008Co-Authors: L V Matasova, T N PopovaAbstract:Data on the structure, functions, regulation of activity, and expression of cytosolic and mitochondrial Aconitate Hydratase isoenzymes of mammals are reviewed. The role of Aconitate Hydratase and structurally similar iron-regulatory protein in maintenance of homeostasis of cell iron is described. Information on modifications of the Aconitate Hydratase molecule and changes in expression under oxidative stress is generalized. The role of Aconitate Hydratase in the pathogenesis of some diseases is considered.
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effects of lipoic acid on citrate content Aconitate Hydratase activity and oxidative status during myocardial ischemia in rats
Biochemistry, 2008Co-Authors: A V Makeeva, T N Popova, L V Matasova, I N YamaAbstract:The effects of lipoic acid on intensity of free radical reactions, citrate content, and Aconitate Hydratase during myocardial ischemia have been investigated. Treatment with lipoic acid normalized biochemiluminescence parameters and citrate level, which were increased in the myocardial pathology. Treatment with lipoic acid also increased specific activity of Aconitate Hydratase, which was decreased in myocardium and blood of animals with myocardial ischemia. Administration of lipoic acid decreased DNA fragmentation observed during myocardial ischemia. The data suggest that lipoic acid can be effectively used as a cardioprotector preventing the development of free radical oxidation during myocardial ischemia.
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the oxidative effects of melatonin the content of citrate and the activity of Aconitate Hydratase in the rat liver in toxic hepatitis
Problemy e̊ndokrinologii, 2005Co-Authors: A N Pashkov, S S Popov, L V Matasova, A V Semenikhina, T N PopovaAbstract:When melatonin was injected into the liver of rats with toxic hepatitis, there was a reduction in the rate of free radical processes and in the mobilization of the antioxidative system, as estimated by the parameters of biochemiluminescence and the level of α-tocopherol, which is likely to be accounted for by the antioxidative effect of this hormone. The administration of melatonin to intact animals causes a reduction in the activity of Aconitate Hydratase and an increase in the level of citrate, which reflects the enhancement of the antioxidative potential as it promotes the decreased hydroxyl radical formation in the Fenton reaction. Metatonin-induced elevations in citrate levels and changes in the activity of Aconitate Hydratase were observed in the liver of animals with toxic hepatitis as compared with those not receiving the hormone, which seems to be associated with the less mobilization of the antioxidative system in the presence of melatonin that is able to produce an antioxidative effect.
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free radical oxidation and catalytic activity of Aconitate Hydratase in rat liver under normal conditions and during toxic hepatitis
Bulletin of Experimental Biology and Medicine, 2004Co-Authors: E M Andreeshcheva, T N Popova, V G Artyukhov, L V MatasovaAbstract:We observed intensification of free radical oxidation, decrease in activity, and changes in catalytic properties of Aconitate Hydratase in the liver of rats with toxic hepatitis. The total yield and maximum flash intensity of biochemiluminescence increased by 2.2 and 1.7 times, respectively. Differences were revealed in the regulation of Aconitate Hydratase activity with Fe2+, Ca2+, H2O2, and oxidized and reduced glutathione in the liver of rats with toxic hepatitis and control animals.
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catalytic properties of cytoplasmic and mitochondrial Aconitate Hydratase from rat cardiomyocytes
Biology Bulletin, 2002Co-Authors: L V Medvedeva, T N Popova, V G Artiukhov, L V MatasovaAbstract:Enzymatic activity of Aconitate Hydratase (aconitase, EC 4.2.1.3) from the rat heart is localized in the cytoplasm (65%) and mitochondria (35%). Cytoplasmic and mitochondrial forms of Aconitate Hydratase were separated by ion-exchange chromatography on CM-Cellulose and CM-Sephadex. The two forms have similar molecular weight, optimal pH range, and spectral properties; however, they have different chromatography properties, Km for citrate and isocitrate, as well as sensitivity to Fe2+ ions.
Manuel Cercos - One of the best experts on this subject based on the ideXlab platform.
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RESEARCH ARTICLE Open Access The Aconitate Hydratase family from Citrus
2013Co-Authors: Javier Terol, Guillermo Soler, Manuel Talon, Manuel CercosAbstract:Background: Research on citrus fruit ripening has received considerable attention because of the importance of citrus fruits for the human diet. Organic acids are among the main determinants of taste and organoleptic quality of fruits and hence the control of fruit acidity loss has a strong economical relevance. In citrus, organic acids accumulate in the juice sac cells of developing fruits and are catabolized thereafter during ripening. Aconitase, that transforms citrate to isocitrate, is the first step of citric acid catabolism and a major component of the citrate utilization machinery. In this work, the citrus aconitase gene family was first characterized and a phylogenetic analysis was then carried out in order to understand the evolutionary history of this family in plants. Gene expression analyses of the citrus aconitase family were subsequently performed in several acidic and acidless genotypes to elucidate their involvement in acid homeostasis. Results: Analysis of 460,000 citrus ESTs, followed by sequencing of complete cDNA clones, identified in citrus 3 transcription units coding for putatively active Aconitate Hydratase proteins, named as CcAco1, CcAco2 and CcAco3. A phylogenetic study carried on the Aco family in 14 plant species, shows the presence of 5 Aco subfamilies, and that the ancestor of monocot and dicot species shared at least one Aco gene. Real-time RT-PCR expression analyses of the three aconitase citrus genes were performed in pulp tissues along fruit development in acidic an
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the Aconitate Hydratase family from citrus
BMC Plant Biology, 2010Co-Authors: Javier Terol, Guillermo Soler, Manuel Talon, Manuel CercosAbstract:Research on citrus fruit ripening has received considerable attention because of the importance of citrus fruits for the human diet. Organic acids are among the main determinants of taste and organoleptic quality of fruits and hence the control of fruit acidity loss has a strong economical relevance. In citrus, organic acids accumulate in the juice sac cells of developing fruits and are catabolized thereafter during ripening. Aconitase, that transforms citrate to isocitrate, is the first step of citric acid catabolism and a major component of the citrate utilization machinery. In this work, the citrus aconitase gene family was first characterized and a phylogenetic analysis was then carried out in order to understand the evolutionary history of this family in plants. Gene expression analyses of the citrus aconitase family were subsequently performed in several acidic and acidless genotypes to elucidate their involvement in acid homeostasis. Analysis of 460,000 citrus ESTs, followed by sequencing of complete cDNA clones, identified in citrus 3 transcription units coding for putatively active Aconitate Hydratase proteins, named as CcAco1, CcAco2 and CcAco3. A phylogenetic study carried on the Aco family in 14 plant species, shows the presence of 5 Aco subfamilies, and that the ancestor of monocot and dicot species shared at least one Aco gene. Real-time RT-PCR expression analyses of the three aconitase citrus genes were performed in pulp tissues along fruit development in acidic and acidless citrus varieties such as mandarins, oranges and lemons. While CcAco3 expression was always low, CcAco1 and CcAco2 genes were generally induced during the rapid phase of fruit growth along with the maximum in acidity and the beginning of the acid reduction. Two exceptions to this general pattern were found: 1) Clemenules mandarin failed inducing CcAco2 although acid levels were rapidly reduced; and 2) the acidless "Sucrena" orange showed unusually high levels of expression of both aconitases, an observation correlating with the acidless phenotype. However, in the acidless "Dulce" lemon aconitase expression was normal suggesting that the acidless trait in this variety is not dependent upon aconitases. Phylogenetic studies showed the occurrence of five different subfamilies of Aconitate Hydratase in plants and sequence analyses indentified three active genes in citrus. The pattern of expression of two of these genes, CcAco1 and CcAco2, was normally associated with the timing of acid content reduction in most genotypes. Two exceptions to this general observation suggest the occurrence of additional regulatory steps of citrate homeostasis in citrus.
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The Aconitate Hydratase family from Citrus
BMC Plant Biology, 2010Co-Authors: Javier Terol, Guillermo Soler, Manuel Talon, Manuel CercosAbstract:Background Research on citrus fruit ripening has received considerable attention because of the importance of citrus fruits for the human diet. Organic acids are among the main determinants of taste and organoleptic quality of fruits and hence the control of fruit acidity loss has a strong economical relevance. In citrus, organic acids accumulate in the juice sac cells of developing fruits and are catabolized thereafter during ripening. Aconitase, that transforms citrate to isocitrate, is the first step of citric acid catabolism and a major component of the citrate utilization machinery. In this work, the citrus aconitase gene family was first characterized and a phylogenetic analysis was then carried out in order to understand the evolutionary history of this family in plants. Gene expression analyses of the citrus aconitase family were subsequently performed in several acidic and acidless genotypes to elucidate their involvement in acid homeostasis. Results Analysis of 460,000 citrus ESTs, followed by sequencing of complete cDNA clones, identified in citrus 3 transcription units coding for putatively active Aconitate Hydratase proteins, named as CcAco1 , CcAco2 and CcAco3 . A phylogenetic study carried on the Aco family in 14 plant species, shows the presence of 5 Aco subfamilies, and that the ancestor of monocot and dicot species shared at least one Aco gene. Real-time RT-PCR expression analyses of the three aconitase citrus genes were performed in pulp tissues along fruit development in acidic and acidless citrus varieties such as mandarins, oranges and lemons. While CcAco3 expression was always low, CcAco1 and CcAco2 genes were generally induced during the rapid phase of fruit growth along with the maximum in acidity and the beginning of the acid reduction. Two exceptions to this general pattern were found: 1) Clemenules mandarin failed inducing CcAco2 although acid levels were rapidly reduced; and 2) the acidless "Sucreña" orange showed unusually high levels of expression of both aconitases, an observation correlating with the acidless phenotype. However, in the acidless "Dulce" lemon aconitase expression was normal suggesting that the acidless trait in this variety is not dependent upon aconitases. Conclusions Phylogenetic studies showed the occurrence of five different subfamilies of Aconitate Hydratase in plants and sequence analyses indentified three active genes in citrus. The pattern of expression of two of these genes, CcAco1 and CcAco2 , was normally associated with the timing of acid content reduction in most genotypes. Two exceptions to this general observation suggest the occurrence of additional regulatory steps of citrate homeostasis in citrus.
