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2. Structure and Biology of Self Lipid Antigens

Author

De Libero, G., Mori, L., Compans, R. W., editor, Cooper, M. D., editor, Honjo, T., editor, Koprowski, H., editor, Melchers, F., editor, Oldstone, M. B. A., editor, Olsnes, S., editor, Svanborg, C., editor, Vogt, P. K., editor, Wagner, H., editor, and Moody, D. Branch, editor

Published
2007
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3. Human T cells engineered with a leukemia lipid-specific TCR enables donor-unrestricted recognition of CD1c-expressing leukemia

Author

Consonni, M, Garavaglia, C, Grilli, A, de Lalla, C, Mancino, A, Mori, L, De Libero, G, Montagna, D, Casucci, M, Serafini, M, Bonini, C, Häussinger, D, Ciceri, F, Bernardi, M, Mastaglio, S, Bicciato, S, Dellabona, P, Casorati, G, Consonni M, Garavaglia C, Grilli A, de Lalla C, Mancino A, Mori L, De Libero G, Montagna D, Casucci M, Serafini M, Bonini C, Häussinger D, Ciceri F, Bernardi M, Mastaglio S, Bicciato S, Dellabona P, Casorati G, Consonni, M, Garavaglia, C, Grilli, A, de Lalla, C, Mancino, A, Mori, L, De Libero, G, Montagna, D, Casucci, M, Serafini, M, Bonini, C, Häussinger, D, Ciceri, F, Bernardi, M, Mastaglio, S, Bicciato, S, Dellabona, P, Casorati, G, Consonni M, Garavaglia C, Grilli A, de Lalla C, Mancino A, Mori L, De Libero G, Montagna D, Casucci M, Serafini M, Bonini C, Häussinger D, Ciceri F, Bernardi M, Mastaglio S, Bicciato S, Dellabona P, and Casorati G

Abstract

Acute leukemia relapsing after chemotherapy plus allogeneic hematopoietic stem cell transplantation can be treated with donor-derived T cells, but this is hampered by the need for donor/recipient MHC-matching and often results in graft-versus-host disease, prompting the search for new donor-unrestricted strategies targeting malignant cells. Leukemia blasts express CD1c antigen-presenting molecules, which are identical in all individuals and expressed only by mature leukocytes, and are recognized by T cell clones specific for the CD1c-restricted leukemia-associated methyl-lysophosphatidic acid (mLPA) lipid antigen. Here, we show that human T cells engineered to express an mLPA-specific TCR, target diverse CD1c-expressing leukemia blasts in vitro and significantly delay the progression of three models of leukemia xenograft in NSG mice, an effect that is boosted by mLPA-cellular immunization. These results highlight a strategy to redirect T cells against leukemia via transfer of a lipid-specific TCR that could be used across MHC barriers with reduced risk of graft-versus-host disease.

Published
2021

5. Aplicación de terapia de presión negativa en el manejo de pacientes con heridas complejas

Author

Barbara Goyo N., Miriamgeluis Lanzotti S., Aracelys Torrealba A., and Libero G De Felice

Subjects
lcsh:R, lcsh:Medicine, presión negativa, heridas complejas, lcsh:Q, vac, lcsh:Science
Abstract

Una herida compleja es aquella que no cicatriza a la velocidad esperada ni con el tratamiento convencional o que recurre una vez cicatrizada. La siguiente investigación se plantea como objetivo aplicar la terapia de presión negativa en el manejo de pacientes con heridas complejas. Materiales y Métodos: Estudio de cohorte observacional, incluyó nueve pacientes con heridas complejas que ingresaron al Hospital Lic. José María Benítez en el periodo comprendido entre febrero-julio de 2019. De los individuos estudiados seis fueron masculinos y tres femeninos, con una media de edad de 50 años, comorbilidades asociadas desnutrición y diabetes mellitus; en cuanto al tipo de herida según el diagrama de C. Leal, la más frecuente fue tipo 4 (55,6%), con diámetro de 11 a 20cm, una profundidad >3 cm, y presencia de abundante exudado purulento; los gérmenes causales más comunes cultivados fueron E.coli (44,4%) y S. aureus (44,4%); En la totalidad de los individuos estudiados se observó la mejoría clínica de la herida en todas sus características, y la iniciación en tres días promedio del tejido de granulación. Concluyendo que con la aplicación del método en estudio hubo aceleración del proceso de cicatrización, control de la infección y disminución de la estancia hospitalaria.

Published
2020

6. Correlation Between TCRV Gene Usage and Antigen Specificities in Human γδ T Cells

Author

De Libero, G., Casorati, G., Migone, N., Lanzavecchia, A., Compans, R. W., editor, Cooper, M., editor, Koprowski, H., editor, McConnell, I., editor, Melchers, F., editor, Nussenzweig, V., editor, Oldstone, M., editor, Olsnes, S., editor, Potter, M., editor, Saedler, H., editor, Vogt, P. K., editor, Wagner, H., editor, Wilson, I., editor, Pfeffer, Klaus, editor, Heeg, Klaus, editor, Wagner, Hermann, editor, and Riethmüller, Gert, editor

Published
1991
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9. Dynamic mode decomposition of GRACE satellite data.

Author

Libero, G., Ciriello, V., and Tartakovsky, D.M.

Subjects
*REDUCED-order models, *DATA compression, *TIME series analysis, *HYDROLOGIC models, *SYSTEM dynamics, *WATER storage
Abstract

Advancements in satellite technology yield environmental data with ever improving spatial coverage and temporal resolution. This necessitates the development of techniques to discern actionable information from large amounts of such data. We explore the potential of dynamic mode decomposition (DMD) to discover the dynamics of spatially correlated structures present in global-scale data, specifically in observations of total water storage anomalies provided by GRACE satellite missions. Our results demonstrate that DMD enables data compression and extrapolation from a reduced set of dominant spatiotemporal structures. The accuracy of its predictions of global system dynamics is preserved in its reconstruction of local time series. These findings suggest potential uses of DMD in analysis of remote-sensing data for hydrologic applications. • DMD accuracy in GRACE mission data representation and extrapolation is quantified at the global scale. • The error of DMD-based reduced-order models (ROMs) is assessed at different latitudes and longitudes. • DMD of global data provides accurate reconstruction of local time series. [ABSTRACT FROM AUTHOR]

Published
2024
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13. Aplicación de terapia de presión negativa en el manejo de pacientes con heridas complejas

Author

Goyo N., Barbara, Lanzotti S., Miriamgeluis, Torrealba A., Aracelys, De Felice, Libero G, Goyo N., Barbara, Lanzotti S., Miriamgeluis, Torrealba A., Aracelys, and De Felice, Libero G

Abstract

A complex wound is the type of wound that has a slow healing process even with standard treatments or tends to recur once it was healed. The aim of this research is to apply negative-pressure wound therapy in the treatment of acute conflict-related wound patients. Materials and Methods: The cohort study included nine patients with complex wounds who were admitted to Hospital Lic. José María Benítez from February to July 2019. Six of the patients on study were male and three female patients whose average age was 50, with comorbidities associated to malnourishment and diabetes mellitus. According to diagram C. Leal, the type of wound mostly found was type 4 (55.6%), ranging from 11-20 cm in diameter, >3 cm depth, with excessive purulent exudate (drainage). The most common cultivated germs found were E. coli (44.4%) and S. aureus (44.4%). All of the patients on study reported clinical improvement of their wounds in all aspects, and an evidence of a 3-day average for the development of the granulation tissue. Concluding that the applying this method evidenced acceleration of wound healing, control of the risk of infection, and reduction of the length of stay in the hospital., Una herida compleja es aquella que no cicatriza a la velocidad esperada ni con el tratamiento convencional o que recurre una vez cicatrizada. La siguiente investigación se plantea como objetivo aplicar la terapia de presión negativa en el manejo de pacientes con heridas complejas. Materiales y Métodos: Estudio de cohorte observacional, incluyó nueve pacientes con heridas complejas que ingresaron al Hospital Lic. José María Benítez en el periodo comprendido entre febrero-julio de 2019. De los individuos estudiados seis fueron masculinos y tres femeninos, con una media de edad de 50 años, comorbilidades asociadas desnutrición y diabetes mellitus; en cuanto al tipo de herida según el diagrama de C. Leal, la más frecuente fue tipo 4 (55,6%), con diámetro de 11 a 20cm, una profundidad >3 cm, y presencia de abundante exudado purulento; los gérmenes causales más comunes cultivados fueron E.coli (44,4%) y S. aureus (44,4%); En la totalidad de los individuos estudiados se observó la mejoría clínica de la herida en todas sus características, y la iniciación en tres días promedio del tejido de granulación.Concluyendo que con la aplicación del método en estudio hubo aceleración del proceso de cicatrización, control de la infección y disminución de la estancia hospitalaria.

Published
2020

18. Peroxisome-derived lipids are self antigens that stimulate invariant natural killer T cells in the thymus

Author

Facciotti, F, Ramanjaneyulu, G, Lepore, M, Sansano, S, Cavallari, M, Kistowska, M, Forss-Petter, S, Ni, G, Colone, A, Singhal, A, Berger, J, Xia, C, Mori, L, De Libero, G, Facciotti F, Ramanjaneyulu GS, Lepore M, Sansano S, Cavallari M, Kistowska M, Forss-Petter S, Ni GH, Colone A, Singhal A, Berger J, Xia CF, Mori L, De Libero G, Facciotti, F, Ramanjaneyulu, G, Lepore, M, Sansano, S, Cavallari, M, Kistowska, M, Forss-Petter, S, Ni, G, Colone, A, Singhal, A, Berger, J, Xia, C, Mori, L, De Libero, G, Facciotti F, Ramanjaneyulu GS, Lepore M, Sansano S, Cavallari M, Kistowska M, Forss-Petter S, Ni GH, Colone A, Singhal A, Berger J, Xia CF, Mori L, and De Libero G

Abstract

The development and maturation of semi-invariant natural killer T cells (iNKT cells) rely on the recognition of self antigens presented by CD1d restriction molecules in thymus. The nature of the stimulatory thymic self lipids remains elusive. We isolated lipids from thymocytes and found that ether-bonded mono-alkyl glycerophosphates and the precursors and degradation products of plasmalogens stimulated iNKT cells. Synthetic analogs showed high potency in activating thymic and peripheral iNKT cells. Mice deficient in the peroxisomal enzyme glyceronephosphate O-acyltransferase (GNPAT), essential for the synthesis of ether lipids, had significant alteration of the thymic maturation of iNKT cells and fewer iNKT cells in both thymus and peripheral organs, which confirmed the role of ether-bonded lipids as iNKT cell antigens. Thus, peroxisome-derived lipids are nonredundant self antigens required for the generation of a full iNKT cell repertoire.

Published
2012

19. Fine tuning by human CD1e of lipid-specific immune responses

Author

Facciotti, F, Cavallari, M, Angenieux, C, Garcia-Alles, L, Signorino-Gelo, F, Angman, L, Gilleron, M, Prandi, J, Puzo, G, Panza, L, Xia, C, Wang, P, Dellabona, P, Casorati, G, Porcelli, S, de la Salle, H, Mori, L, De Libero, G, Facciotti F, Cavallari M, Angenieux C, Garcia-Alles LF, Signorino-Gelo F, Angman L, Gilleron M, Prandi J, Puzo G, Panza L, Xia CF, Wang PG, Dellabona P, Casorati G, Porcelli SA, de la Salle H, Mori L, De Libero G, Facciotti, F, Cavallari, M, Angenieux, C, Garcia-Alles, L, Signorino-Gelo, F, Angman, L, Gilleron, M, Prandi, J, Puzo, G, Panza, L, Xia, C, Wang, P, Dellabona, P, Casorati, G, Porcelli, S, de la Salle, H, Mori, L, De Libero, G, Facciotti F, Cavallari M, Angenieux C, Garcia-Alles LF, Signorino-Gelo F, Angman L, Gilleron M, Prandi J, Puzo G, Panza L, Xia CF, Wang PG, Dellabona P, Casorati G, Porcelli SA, de la Salle H, Mori L, and De Libero G

Abstract

CD1e is a member of the CD1 family that participates in lipid antigen presentation without interacting with the T-cell receptor. It binds lipids in lysosomes and facilitates processing of complex glycolipids, thus promoting editing of lipid antigens. We find that CD1e may positively or negatively affect lipid presentation by CD1b, CD1c, and CD1d. This effect is caused by the capacity of CD1e to facilitate rapid formation of CD1-lipid complexes, as shown for CD1d, and also to accelerate their turnover. Similar results were obtained with antigen-presenting cells from CD1e transgenic mice in which lipid complexes are assembled more efficiently and show faster turnover than in WT antigen-presenting cells. These effects maximize and temporally narrow CD1-restricted responses, as shown by reactivity to Sphingomonas paucimobilis-derived lipid antigens. CD1e is therefore an important modulator of both group 1 and group 2 CD1-restricted responses influencing the lipid antigen availability as well as the generation and persistence of CD1-lipid complexes.

Published
2011

20. Thymocytes control the CD4 gene differently from mature T lymphocytes

Author

Uematsu, Y., Donda, A., De Libero, G., Uematsu, Y., Donda, A., and De Libero, G.

Abstract

We analyzed the activity of the enhancer, the promoter and the silencer of the human CD4 gene during T cell development using transgenic mice. Immunofluorescence studies on thymic populations of mice carrying transgenes in various combinations of these regulatory DNA elements revealed that thymocytes control the CD4 gene in a different manner than mature peripheral T lymphocytes. The 5'-positive regulatory unit, consisting of the promoter and the 5' enhancer, is already active at the CD4-CD8-double-negative (DN) stage of development. However, its activity becomes lower in the double-positive and a fraction of the CD4+ CD8int/- cell population, indicating that an additional enhancer, located in either the first or the third intron of the CD4 gene, is required for CD4 gene expression in this population. The other studied regulatory element is the minimal CD4 silencer which inhibits CD4 gene expression in peripheral CD8 T lymphocytes. This silencer is inactive in the most immature DN thymocytes, which probably use a distinct silencer mechanism to down-regulate CD4 gene expression. Unexpectedly, the CD4 silencer is also active in CD4+ CD8int/- cells of the thymus, implying that an anti-silencer may be required to resume CD4 expression in this cell population. Altogether, the CD4 gene is regulated by several positive and negative regulatory mechanisms which come into play in a developmentally coordinated manner

Published
2017

21. T cell epitope-mapping by cytokine gene expression assay

Author

De Libero, G, Walker, J M, De Libero, G ( G ), Walker, J M ( J M ), Provenzano, M; https://orcid.org/0000-0001-6993-5718, Spagnoli, G C, De Libero, G, Walker, J M, De Libero, G ( G ), Walker, J M ( J M ), Provenzano, M; https://orcid.org/0000-0001-6993-5718, and Spagnoli, G C

Abstract

The following method describes the identification of candidate immunogenic peptides through their ability to recall an immune T-cell activation from peripheral blood mononuclear cells (PBMCs) of individuals with defined HLA-peptide restrictions that have been previously exposed to the antigen. Isolated PBMCs are plated out at a concentration of 1 x 10(6) cells/ml in a 200 microl medium and incubated overnight to reduce cytokine gene expression due to cell manipulation. After starving, cells are either directly stimulated with individual peptides or not stimulated and incubated from 3 to 12 h according to experimental conditions. Quantitative real-time PCR (qrt-PCR) is performed on reverse-transcribed complementary DNA (cDNA) from total RNA that is isolated from peptide-cultured PBMCs. To perform high quality qrt-PCR, primers and probes are designed to span exon-intron junctions in order to prevent amplification of genomic DNA and to produce amplicons <150 base pairs (bp). Real-time monitoring of fluorescent emission from the cleavage of sequence specific probe by the nuclease activity of Taq polymerase (TaqMan method) defines threshold cycles during exponential phases of amplification. Standard curves of copy numbers of the gene of interest are normalized using as reference copy numbers of control genes.

Published
2009

22. Lipids from leukemia cells stimulate CD1c-reactive T cells

Author

Lepore M, Gundimeda SR, Sansano S, Mori L, De Libero G, De Lalla C, Dellabona P, Casorati G, Montagna D, CICERI , FABIO, Lepore, M, Gundimeda, Sr, Sansano, S, Mori, L, De Libero, G, De Lalla, C, Dellabona, P, Casorati, G, Montagna, D, and Ciceri, Fabio

Published
2011

23. Differential alteration of lipid antigen presentation to NKT cells due to imbalances in lipid metabolism

Author

Schumann, J, Facciotti, F, Panza, L, Michieletti, M, Compostella, F, Collmann, A, Mori, L, De Libero, G, Schumann J, Facciotti F, Panza L, Michieletti M, Compostella F, Collmann A, Mori L, De Libero G, Schumann, J, Facciotti, F, Panza, L, Michieletti, M, Compostella, F, Collmann, A, Mori, L, De Libero, G, Schumann J, Facciotti F, Panza L, Michieletti M, Compostella F, Collmann A, Mori L, and De Libero G

Abstract

Deficiencies in enzymes of the lysosomal glycosphingolipid degradation pathway or in lysosomal lipid transfer proteins cause an imbalance in lipid metabolism and induce accumulation of certain lipids. A possible impact of such an imbalance on the presentation of lipid antigens to lipid-reactive T cells has only been hypothesized but not extensively studied so far. Here we demonstrate that presentation of lipid antigens to, and development of, lipid-reactive CD1d-restricted NKT cells, are impaired in mice deficient in the lysosomal enzyme P-galactosidase (OGal) or the lysosomal lipid transfer protein Niemann-Pick C (NPC) 2. Importantly, the residual populations of NKT cells selected in beta Gal(-/-) and NPC2(-/-) mice showed differential TCR and CD4 repertoire characteristics, suggesting that differential selecting CD1d:lipid antigen complexes are formed. Furthermore, we provide direct evidence that accumulation of lipids impairs lipid antigen presentation in both cases. However, the mechanisms by which imbalanced lipid metabolism affected lipid antigen presentation were different. Based on these results, the impact of lipid accumulation should be generally considered in the interpretation of immunological deficiencies found in mice suffering from lipid metabolic disorders.

Published
2007

24. A general and stereoselective route to alpha- or beta-galactosphingolipids via a common four-carbon building block

Author

Matto, P, Modica, E, Franchini, L, Facciotti, F, Mori, L, De Libero, G, Lombardi, G, Fallarini, S, Panza, L, Compostella, F, Ronchetti, F, Matto P, Modica E, Franchini L, Facciotti F, Mori L, De Libero G, Lombardi G, Fallarini S, Panza L, Compostella F, Ronchetti F, Matto, P, Modica, E, Franchini, L, Facciotti, F, Mori, L, De Libero, G, Lombardi, G, Fallarini, S, Panza, L, Compostella, F, Ronchetti, F, Matto P, Modica E, Franchini L, Facciotti F, Mori L, De Libero G, Lombardi G, Fallarini S, Panza L, Compostella F, and Ronchetti F

Abstract

A general synthetic strategy toward alpha- or beta-galactosylceramides and their analogues from 3-azido-2-O-benzyl-1-O-(4-methoxybenzyl)butane- 1,2,4-triol is described. The key steps for the installation of the main lipid chain are either a diasteroselective alkynylation reaction yielding the 4R stereocenter of phytosphingosine or a Wittig olefination generating the trans double bond of sphingosine. The methodology allows the preparation of different glycolipids with variations in the structure of the sphingoid base. In particular, three alpha-GalCer-related compounds have been synthesized and evaluated for their ability to activate CD1d-restricted T-cells.

Published
2007

25. Ribulose-1,5-bisphosphate regeneration in the Calvin-Benson-Bassham cycle: Focus on the last three enzymatic steps that allow the formation of Rubisco substrate

Author

Maria Meloni, Libero Gurrieri, Simona Fermani, Lauren Velie, Francesca Sparla, Pierre Crozet, Julien Henri, and Mirko Zaffagnini

Subjects
carbon fixation, structure, catalysis, isomerase, epimerase, kinase, Plant culture, SB1-1110
Abstract

The Calvin-Benson-Bassham (CBB) cycle comprises the metabolic phase of photosynthesis and is responsible for carbon fixation and the production of sugar phosphates. The first step of the cycle involves the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) which catalyzes the incorporation of inorganic carbon into 3-phosphoglyceric acid (3PGA). The following steps include ten enzymes that catalyze the regeneration of ribulose-1,5-bisphosphate (RuBP), the substrate of Rubisco. While it is well established that Rubisco activity acts as a limiting step of the cycle, recent modeling studies and experimental evidence have shown that the efficiency of the pathway is also impacted by the regeneration of the Rubisco substrate itself. In this work, we review the current understanding of the structural and catalytic features of the photosynthetic enzymes that catalyze the last three steps of the regeneration phase, namely ribose-5-phosphate isomerase (RPI), ribulose-5-phosphate epimerase (RPE), and phosphoribulokinase (PRK). In addition, the redox- and metabolic-based regulatory mechanisms targeting the three enzymes are also discussed. Overall, this review highlights the importance of understudied steps in the CBB cycle and provides direction for future research aimed at improving plant productivity.

Published
2023
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26. Polysaccharide antigen-glycolipid conjugate vaccines

Author

Seeberger, P., Stallforth, P., De Libero, G., and Cavallari, M.

Abstract

The present invention relates to the field of synthesizing and biologically evaluating of a novel class of carbohydratebased vaccines. The new vaccines consist of a multi- modular structure which allows applying the vaccine to a whole variety of pathogenes. This method allows preparing vaccines against all pathogens expressing immunogenic carbohydrate antigens. As conjugation of antigenic carbohydrates to proteins is not required the conjugate vaccine is particularly heat stable. No refrigeration is required, a major drawback of protein-based vaccines.

Published
2013

28. Proline, Cysteine and Branched-Chain Amino Acids in Abiotic Stress Response of Land Plants and Microalgae

Author

Rachele Ingrisano, Edoardo Tosato, Paolo Trost, Libero Gurrieri, and Francesca Sparla

Subjects
amino acids, abiotic stress, proline, cysteine, hydrogen sulfide, catabolism, Botany, QK1-989
Abstract

Proteinogenic amino acids are the building blocks of protein, and plants synthesize all of them. In addition to their importance in plant growth and development, growing evidence underlines the central role played by amino acids and their derivatives in regulating several pathways involved in biotic and abiotic stress responses. In the present review, we illustrate (i) the role of amino acids as an energy source capable of replacing sugars as electron donors to the mitochondrial electron transport chain and (ii) the role of amino acids as precursors of osmolytes as well as (iii) precursors of secondary metabolites. Among the amino acids involved in drought stress response, proline and cysteine play a special role. Besides the large proline accumulation occurring in response to drought stress, proline can export reducing equivalents to sink tissues and organs, and the production of H2S deriving from the metabolism of cysteine can mediate post-translational modifications that target protein cysteines themselves. Although our general understanding of microalgae stress physiology is still fragmentary, a general overview of how unicellular photosynthetic organisms deal with salt stress is also provided because of the growing interest in microalgae in applied sciences.

Published
2023
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31. CD1e participates in processing of microbial glycolipid antigens

Author

La Salle H., De, Mariotti, S., Angenieux, C., Gilleron, M., L.-F., Garcia-Alles, Maître, B., Malm, D., Berg, T., Mourey, L., Salamero, J., J.P., Cazenave, Hanau, D., Mori, L., Puzo, G., Libero G., De, Institut de pharmacologie et de biologie structurale (IPBS), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Published
2005

33. Arabidopsis thaliana Sucrose Phosphate Synthase A2 Affects Carbon Partitioning and Drought Response

Author

Laura Bagnato, Edoardo Tosato, Libero Gurrieri, Paolo Trost, Giuseppe Forlani, and Francesca Sparla

Subjects
osmotic stress, sucrose phosphate synthase, glucose-6-phosphate dehydrogenase, fructose, oxidative pentose phosphate pathway, Biology (General), QH301-705.5
Abstract

Sucrose is essential for plants for several reasons: It is a source of energy, a signaling molecule, and a source of carbon skeletons. Sucrose phosphate synthase (SPS) catalyzes the conversion of uridine diphosphate glucose and fructose-6-phosphate to sucrose-6-phosphate, which is rapidly dephosphorylated by sucrose phosphatase. SPS is critical in the accumulation of sucrose because it catalyzes an irreversible reaction. In Arabidopsis thaliana, SPSs form a gene family of four members, whose specific functions are not clear yet. In the present work, the role of SPSA2 was investigated in Arabidopsis under both control and drought stress conditions. In seeds and seedlings, major phenotypic traits were not different in wild-type compared with spsa2 knockout plants. By contrast, 35-day-old plants showed some differences in metabolites and enzyme activities even under control conditions. In response to drought, SPSA2 was transcriptionally activated, and the divergences between the two genotypes were higher, with spsa2 showing reduced proline accumulation and increased lipid peroxidation. Total soluble sugars and fructose concentrations were about halved compared with wild-type plants, and the plastid component of the oxidative pentose phosphate pathway was activated. Unlike previous reports, our results support the involvement of SPSA2 in both carbon partitioning and drought response.

Published
2023
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36. Endogenous phosphatidylcholine and an long spacer ligand stabilize the lipid-binding groove of CD1b

Author

Biomoleculaire Massaspectrometrie, Massaspectrometrie, Dep Scheikunde, Garcia-Alles, L.F., Versluis, C., Maveyraud, L., Vallina, A.T., Sansano, S., Bello, N.F., Gober, H-J, Guillet, V., De la Salle, H., Puzo, G., Mori, L., Heck, A.J.R., De Libero, G., Mourey, L., Biomoleculaire Massaspectrometrie, Massaspectrometrie, Dep Scheikunde, Garcia-Alles, L.F., Versluis, C., Maveyraud, L., Vallina, A.T., Sansano, S., Bello, N.F., Gober, H-J, Guillet, V., De la Salle, H., Puzo, G., Mori, L., Heck, A.J.R., De Libero, G., and Mourey, L.

Published
2006

37. An alternative approach to the assessment of gamma delta T-cell clonality in celiac disease intestinal lesions through cDNA heteroduplex analysis of T-cell receptor VJ junctions

Author

Claudia Giachino, Nicola Migone, N. Ansaldi, De Libero G, Rocci Mp, and Giuseppina Oderda

Subjects
Adult, Male, Adolescent, T-Lymphocytes, Molecular Sequence Data, Immunology, Biology, Polymerase Chain Reaction, Intestinal mucosa, Complementary DNA, medicine, Humans, Immunology and Allergy, Child, Gamma delta T cell, Cloning, Base Sequence, T-cell receptor, Nucleic Acid Heteroduplexes, Infant, Receptors, Antigen, T-Cell, gamma-delta, General Medicine, Molecular biology, In vitro, Clone Cells, Intestines, Celiac Disease, medicine.anatomical_structure, Polyclonal antibodies, Child, Preschool, biology.protein, Female, Heteroduplex
Abstract

We have investigated the clonality of the γδ T lymphocytes infiltrating the intestinal mucosa of CD patients and control subjects by means of a simple and powerful method based on the heteroduplex analysis of the TCR VJ junctions. Each V-specific TCR chain, amplified either from fresh biopsy material or intestinal T-cell-line cDNA, is denatured and renatured to allow the random reshuffling of the various strands carrying different junctional sequences, coamplified in the same reaction. The mismatched chains (heteroduplexes) are separated from the matched ones (homoduplexes) through polyacrylamide gel electrophoresis, and whenever one or more T-cell clones are emerging over the polyclonal background, discrete bands are visible by ethidium-bromide staining. Through this method, we have estimated the diversity of the Vδ1–3 chains and a newly described V gene (Vδ8) whose homologue in mice is abundantly expressed in γδ iLs. We demonstrate that the well-documented expansion of Vγ1 + γδ lymphocytes in the jejunum of CD patients is polyclonal. Overall, the heteroduplex analysis on fresh intestinal and peripheral blood lymphocytes from both healthy and affected subjects shows a polyclonal pattern of all the Vδ + subsets. In contrast, most intestinal T-cell lines produce oligoclonal patterns, suggesting a dramatic in vitro selection effect. The cell expansion in culture is generally not required for the TCR heteroduplex analysis, which can therefore be applied to rapidly monitor the T-cell response in a variety of physiologic and autoimmune reactions, substituting the standard approach of TCR cloning and multiple VJ sequencing. Human Immunology 40, 303–311 (1994)

Published
1994

38. Saturday, 17 July 2010

Author

Dimova, I., primary, Hlushchuk, R., additional, Makanya, A., additional, Djonov, V., additional, Theurl, M., additional, Schgoer, W., additional, Albrecht, K., additional, Beer, A., additional, Patsch, J. R., additional, Schratzberger, P., additional, Mahata, S., additional, Kirchmair, R., additional, Didie, M., additional, Christalla, P., additional, Rau, T., additional, Eschenhagen, T., additional, Schumacher, U., additional, Lin, Q., additional, Zenke, M., additional, Zimmmermann, W., additional, Hoch, M., additional, Fischer, P., additional, Stapel, B., additional, Missol-Kolka, E., additional, Erschow, S., additional, Scherr, M., additional, Drexler, H., additional, Hilfiker-Kleiner, D., additional, Diebold, I., additional, Petry, A., additional, Kennel, P., additional, Djordjevic, T., additional, Hess, J., additional, Goerlach, A., additional, Castellano, J., additional, Aledo, R., additional, Sendra, J., additional, Costales, P., additional, Badimon, L., additional, Llorente-Cortes, V., additional, Dworatzek, E., additional, Mahmoodzadeh, S., additional, Regitz-Zagrosek, V., additional, Posa, A., additional, Varga, C., additional, Berko, A., additional, Veszelka, M., additional, Szablics, P., additional, Vari, B., additional, Pavo, I., additional, Laszlo, F., additional, Brandenburger, M., additional, Wenzel, J., additional, Bogdan, R., additional, Richardt, D., additional, Reppel, M., additional, Hescheler, J., additional, Terlau, H., additional, Dendorfer, A., additional, Heijman, J., additional, Rudy, Y., additional, Westra, R., additional, Volders, P., additional, Rasmusson, R., additional, Bondarenko, V., additional, Ertas Gokhan, M. D., additional, Ural Ertan, M. D., additional, Karaoz Erdal, P. H. D., additional, Aksoy Ayca, P. H. D., additional, Kilic Teoman, M. D., additional, Kozdag Guliz, M. D., additional, Vural Ahmet, M. D., additional, Ural Dilek, M. D., additional, Poulet, C., additional, Christ, T., additional, Wettwer, E., additional, Ravens, U., additional, Van Der Pouw Kraan, C., additional, Schirmer, S., additional, Fledderus, J., additional, Moerland, P., additional, Leyen, T., additional, Piek, J., additional, Van Royen, N., additional, Horrevoets, A., additional, Fleissner, F., additional, Jazbutyte, V., additional, Fiedler, J., additional, Galuppo, P., additional, Mayr, M., additional, Ertl, G., additional, Bauersachs, J., additional, Thum, T., additional, Protze, S., additional, Bussek, A., additional, Li, F., additional, Hoo, R., additional, Lam, K., additional, Xu, A., additional, Subramanian, P., additional, Karshovska, E., additional, Megens, R., additional, Akhtar, S., additional, Heyll, K., additional, Jansen, Y., additional, Weber, C., additional, Schober, A., additional, Zafeiriou, M., additional, Noack, C., additional, Renger, A., additional, Dietz, R., additional, Zelarayan, L., additional, Bergmann, M., additional, Meln, I., additional, Malashicheva, A., additional, Anisimov, S., additional, Kalinina, N., additional, Sysoeva, V., additional, Zaritskey, A., additional, Barbuti, A., additional, Scavone, A., additional, Mazzocchi, N., additional, Crespi, A., additional, Capilupo, D., additional, Difrancesco, D., additional, Qian, L., additional, Shim, W., additional, Gu, Y., additional, Mohammed, S., additional, Wong, P., additional, Zafiriou, M., additional, Schaeffer, H., additional, Kovacs, P., additional, Simon, J., additional, Varro, A., additional, Athias, P., additional, Wolf, J., additional, Bouchot, O., additional, Vandroux, D., additional, Mathe, A., additional, De Carvalho, A., additional, Laurent, G., additional, Rainer, P., additional, Huber, M., additional, Edelmann, F., additional, Stojakovic, T., additional, Trantina-Yates, A., additional, Trauner, M., additional, Pieske, B., additional, Von Lewinski, D., additional, De Jong, A., additional, Maass, A., additional, Oberdorf-Maass, S., additional, Van Gelder, I., additional, Lin, Y., additional, Li, J., additional, Wang, F., additional, He, Y., additional, Li, X., additional, Xu, H., additional, Yang, X., additional, Coppini, R., additional, Ferrantini, C., additional, Ferrara, C., additional, Rossi, A., additional, Mugelli, A., additional, Poggesi, C., additional, Cerbai, E., additional, Rozmaritsa, N., additional, Voigt, N., additional, Dobrev, D., additional, Kienitz, M.-C., additional, Zoidl, G., additional, Bender, K., additional, Pott, L., additional, Kohajda, Z., additional, Kristof, A., additional, Virag, L., additional, Jost, N., additional, Trafford, A., additional, Prnjavorac, B., additional, Mujaric, E., additional, Jukic, J., additional, Abduzaimovic, K., additional, Brack, K., additional, Patel, V., additional, Coote, J., additional, Ng, G., additional, Wilders, R., additional, Van Ginneken, A., additional, Verkerk, A., additional, Xaplanteris, P., additional, Vlachopoulos, C., additional, Baou, K., additional, Vassiliadou, C., additional, Dima, I., additional, Ioakeimidis, N., additional, Stefanadis, C., additional, Ruifrok, W., additional, Qian, C., additional, Sillje, H., additional, Van Goor, H., additional, Van Veldhuisen, D., additional, Van Gilst, W., additional, De Boer, R., additional, Schmidt, K., additional, Kaiser, F., additional, Erdmann, J., additional, De Wit, C., additional, Barnett, O., additional, Kyyak, Y., additional, Cesana, F., additional, Boffi, L., additional, Mauri, T., additional, Alloni, M., additional, Betelli, M., additional, Nava, S., additional, Giannattasio, C., additional, Mancia, G., additional, Vilskersts, R., additional, Kuka, J., additional, Svalbe, B., additional, Liepinsh, E., additional, Dambrova, M., additional, Zakrzewicz, A., additional, Maroski, J., additional, Vorderwuelbecke, B., additional, Fiedorowicz, K., additional, Da Silva-Azevedo, L., additional, Pries, A., additional, Gryglewska, B., additional, Necki, M., additional, Zelawski, M., additional, Grodzicki, T., additional, Scoditti, E., additional, Massaro, M., additional, Carluccio, M., additional, Distante, A., additional, Storelli, C., additional, De Caterina, R., additional, Kocgirli, O., additional, Valcaccia, S., additional, Dao, V., additional, Suvorava, T., additional, Kumpf, S., additional, Floeren, M., additional, Oppermann, M., additional, Kojda, G., additional, Leo, C., additional, Ziogas, J., additional, Favaloro, J., additional, Woodman, O., additional, Goettsch, W., additional, Marton, A., additional, Goettsch, C., additional, Morawietz, H., additional, Khalifa, E., additional, Ashour, Z., additional, Rupprecht, V., additional, Scalera, F., additional, Martens-Lobenhoffer, J., additional, Bode-Boeger, S., additional, Li, W., additional, Kwan, Y., additional, Leung, G., additional, Patella, F., additional, Mercatanti, A., additional, Pitto, L., additional, Rainaldi, G., additional, Tsimafeyeu, I., additional, Tishova, Y., additional, Wynn, N., additional, Kalinchenko, S., additional, Clemente Lorenzo, M., additional, Grande, M., additional, Barriocanal, F., additional, Aparicio, M., additional, Martin, A., additional, Hernandez, J., additional, Lopez Novoa, J., additional, Martin Luengo, C., additional, Kurlianskaya, A., additional, Denisevich, T., additional, Barth, N., additional, Loot, A., additional, Fleming, I., additional, Wang, Y., additional, Gabrielsen, A., additional, Ripa, R., additional, Jorgensen, E., additional, Kastrup, J., additional, Arderiu, G., additional, Pena, E., additional, Kobus, K., additional, Czyszek, J., additional, Kozlowska-Wiechowska, A., additional, Milkiewicz, P., additional, Milkiewicz, M., additional, Madonna, R., additional, Montebello, E., additional, Geng, Y., additional, Chin-Dusting, J., additional, Michell, D., additional, Skilton, M., additional, Dixon, J., additional, Dart, A., additional, Moore, X., additional, Ehrbar, M., additional, Reichmuth, P., additional, Heinimann, N., additional, Hewing, B., additional, Stangl, V., additional, Stangl, K., additional, Laule, M., additional, Baumann, G., additional, Ludwig, A., additional, Widmer-Teske, R., additional, Mueller, A., additional, Stieger, P., additional, Tillmanns, H., additional, Braun-Dullaeus, R., additional, Sedding, D., additional, Troidl, K., additional, Eller, L., additional, Benli, I., additional, Apfelbeck, H., additional, Schierling, W., additional, Troidl, C., additional, Schaper, W., additional, Schmitz-Rixen, T., additional, Hinkel, R., additional, Trenkwalder, T., additional, Pfosser, A., additional, Globisch, F., additional, Stachel, G., additional, Lebherz, C., additional, Bock-Marquette, I., additional, Kupatt, C., additional, Seyler, C., additional, Duthil-Straub, E., additional, Zitron, E., additional, Scholz, E., additional, Thomas, D., additional, Gierten, J., additional, Karle, C., additional, Fink, R., additional, Padro, T., additional, Lugano, R., additional, Garcia-Arguinzonis, M., additional, Schuchardt, M., additional, Pruefer, J., additional, Toelle, M., additional, Pruefer, N., additional, Jankowski, V., additional, Jankowski, J., additional, Zidek, W., additional, Van Der Giet, M., additional, Fransen, P., additional, Van Hove, C., additional, Michiels, C., additional, Van Langen, J., additional, Bult, H., additional, Quarck, R., additional, Wynants, M., additional, Alfaro-Moreno, E., additional, Rosario Sepulveda, M., additional, Wuytack, F., additional, Van Raemdonck, D., additional, Meyns, B., additional, Delcroix, M., additional, Christofi, F., additional, Wijetunge, S., additional, Sever, P., additional, Hughes, A., additional, Ohanian, J., additional, Forman, S., additional, Ohanian, V., additional, Gibbons, C., additional, Vernia, S., additional, Das, A., additional, Shah, V., additional, Casado, M., additional, Bielenberg, W., additional, Daniel, J., additional, Daniel, J.-M., additional, Hersemeyer, K., additional, Schmidt-Woell, T., additional, Kaetzel, D., additional, Tillmans, H., additional, Kanse, S., additional, Tuncay, E., additional, Kandilci, H., additional, Zeydanli, E., additional, Sozmen, N., additional, Akman, D., additional, Yildirim, S., additional, Turan, B., additional, Nagy, N., additional, Acsai, K., additional, Farkas, A., additional, Papp, J., additional, Toth, A., additional, Viero, C., additional, Mason, S., additional, Williams, A., additional, Marston, S., additional, Stuckey, D., additional, Dyer, E., additional, Song, W., additional, El Kadri, M., additional, Hart, G., additional, Hussain, M., additional, Faltinova, A., additional, Gaburjakova, J., additional, Urbanikova, L., additional, Hajduk, M., additional, Tomaskova, B., additional, Antalik, M., additional, Zahradnikova, A., additional, Steinwascher, P., additional, Jaquet, K., additional, Muegge, A., additional, Wang, G., additional, Zhang, M., additional, Tesi, C., additional, Ter Keurs, H., additional, Kettlewell, S., additional, Smith, G., additional, Workman, A., additional, Lenaerts, I., additional, Holemans, P., additional, Sokolow, S., additional, Schurmans, S., additional, Herchuelz, A., additional, Sipido, K., additional, Antoons, G., additional, Wehrens, X., additional, Li, N., additional, Respress, J. R., additional, De Almeida, A., additional, Van Oort, R., additional, Lohmann, H., additional, Saes, M., additional, Messer, A., additional, Copeland, O., additional, Leung, M., additional, Matthes, F., additional, Steinbrecher, J., additional, Salinas-Riester, G., additional, Opitz, L., additional, Hasenfuss, G., additional, Lehnart, S., additional, Caracciolo, G., additional, Eleid, M., additional, Carerj, S., additional, Chandrasekaran, K., additional, Khandheria, B., additional, Sengupta, P., additional, Riaz, I., additional, Tyng, L., additional, Dou, Y., additional, Seymour, A., additional, Dyer, C., additional, Griffin, S., additional, Haswell, S., additional, Greenman, J., additional, Yasushige, S., additional, Amorim, P., additional, Nguyen, T., additional, Schwarzer, M., additional, Mohr, F., additional, Doenst, T., additional, Popin Sanja, S., additional, Lalosevic, D., additional, Capo, I., additional, Momcilov Popin, T., additional, Astvatsatryan, A., additional, Senan, M., additional, Shafieian, G., additional, Goncalves, N., additional, Falcao-Pires, I., additional, Henriques-Coelho, T., additional, Moreira-Goncalves, D., additional, Leite-Moreira, A., additional, Bronze Carvalho, L., additional, Azevedo, J., additional, Andrade, M., additional, Arroja, I., additional, Relvas, M., additional, Morais, G., additional, Seabra, M., additional, Aleixo, A., additional, Winter, J., additional, Zabunova, M., additional, Mintale, I., additional, Lurina, D., additional, Narbute, I., additional, Zakke, I., additional, Erglis, A., additional, Marcinkevics, Z., additional, Kusnere, S., additional, Abolins, A., additional, Aivars, J., additional, Rubins, U., additional, Nassar, Y., additional, Monsef, D., additional, Hamed, G., additional, Abdelshafy, S., additional, Chen, L., additional, Wu, Y., additional, Wang, J., additional, Cheng, C., additional, Sternak, M., additional, Khomich, T., additional, Jakubowski, A., additional, Szafarz, M., additional, Szczepanski, W., additional, Mateuszuk, L., additional, Szymura-Oleksiak, J., additional, Chlopicki, S., additional, Sulicka, J., additional, Strach, M., additional, Kierzkowska, I., additional, Surdacki, A., additional, Mikolajczyk, T., additional, Balwierz, W., additional, Guzik, T., additional, Dmitriev, V., additional, Oschepkova, E., additional, Polovitkina, O., additional, Titov, V., additional, Rogoza, A., additional, Shakur, R., additional, Metcalfe, S., additional, Bradley, J., additional, Demyanets, S., additional, Kaun, C., additional, Kastl, S., additional, Pfaffenberger, S., additional, Huk, I., additional, Maurer, G., additional, Huber, K., additional, Wojta, J., additional, Eriksson, O., additional, Aberg, M., additional, Siegbahn, A., additional, Niccoli, G., additional, Sgueglia, G., additional, Conte, M., additional, Giubilato, S., additional, Cosentino, N., additional, Ferrante, G., additional, Crea, F., additional, Ilisei, D., additional, Leon, M., additional, Mitu, F., additional, Kyriakakis, E., additional, Philippova, M., additional, Cavallari, M., additional, Bochkov, V., additional, Biedermann, B., additional, De Libero, G., additional, Erne, P., additional, Resink, T., additional, Bakogiannis, C., additional, Antoniades, C., additional, Tousoulis, D., additional, Demosthenous, M., additional, Psarros, C., additional, Sfyras, N., additional, Channon, K., additional, Del Turco, S., additional, Navarra, T., additional, Basta, G., additional, Carnicelli, V., additional, Frascarelli, S., additional, Zucchi, R., additional, Kostareva, A., additional, Sjoberg, G., additional, Gudkova, A., additional, Semernin, E., additional, Shlyakhto, E., additional, Sejersen, T., additional, Cucu, N., additional, Anton, M., additional, Stambuli, D., additional, Botezatu, A., additional, Arsene, C., additional, Lupeanu, E., additional, Anton, G., additional, Patsch, J., additional, Huber, E., additional, Lande, C., additional, Cecchettini, A., additional, Tedeschi, L., additional, Trivella, M., additional, Citti, L., additional, Chen, B., additional, Ma, Y., additional, Yang, Y., additional, Ma, X., additional, Liu, F., additional, Hasanzad, M., additional, Rejali, L., additional, Fathi, M., additional, Minassian, A., additional, Mohammad Hassani, R., additional, Najafi, A., additional, Sarzaeem, M., additional, Sezavar, S., additional, Akhmedov, A., additional, Klingenberg, R., additional, Yonekawa, K., additional, Lohmann, C., additional, Gay, S., additional, Maier, W., additional, Neithard, M., additional, Luescher, T., additional, Xie, X., additional, Fu, Z., additional, Kevorkov, A., additional, Verduci, L., additional, Cremisi, F., additional, Wonnerth, A., additional, Katsaros, K., additional, Zorn, G., additional, Weiss, T., additional, De Rosa, R., additional, Galasso, G., additional, Piscione, F., additional, Santulli, G., additional, Iaccarino, G., additional, Piccolo, R., additional, Luciano, R., additional, Chiariello, M., additional, Szymanski, M., additional, Schoemaker, R., additional, Hillege, H., additional, Rizzo, S., additional, Basso, C., additional, Thiene, G., additional, Valente, M., additional, Rickelt, S., additional, Franke, W., additional, Bartoloni, G., additional, Bianca, S., additional, Giurato, E., additional, Barone, C., additional, Ettore, G., additional, Bianca, I., additional, Eftekhari, P., additional, Wallukat, G., additional, Bekel, A., additional, Heinrich, F., additional, Fu, M., additional, Briedert, M., additional, Briand, J., additional, Roegel, J., additional, Pilichou, K., additional, Korkmaz, S., additional, Radovits, T., additional, Pali, S., additional, Hirschberg, K., additional, Zoellner, S., additional, Loganathan, S., additional, Karck, M., additional, Szabo, G., additional, Pucci, A., additional, Pantaleo, J., additional, Martino, S., additional, Pelosi, G., additional, Matteucci, M., additional, Kusmic, C., additional, Vesentini, N., additional, Piccolomini, F., additional, Viglione, F., additional, L'abbate, A., additional, Slavikova, J., additional, Chottova Dvorakova, M., additional, Kummer, W., additional, Campanile, A., additional, Spinelli, L., additional, Ciccarelli, M., additional, De Gennaro, S., additional, Assante Di Panzillo, E., additional, Trimarco, B., additional, Akbarzadeh Najar, R., additional, Ghaderian, S., additional, Tabatabaei Panah, A., additional, Vakili, H., additional, Rezaei Farimani, A., additional, Rezaie, G., additional, Beigi Harchegani, A., additional, Hamdani, N., additional, Gavina, C., additional, Van Der Velden, J., additional, Niessen, H., additional, Stienen, G., additional, Paulus, W., additional, Moura, C., additional, Lamego, I., additional, Eloy, C., additional, Areias, J., additional, Bonda, T., additional, Dziemidowicz, M., additional, Hirnle, T., additional, Dmitruk, I., additional, Kaminski, K., additional, Musial, W., additional, Winnicka, M., additional, Villar, A., additional, Merino, D., additional, Ares, M., additional, Pilar, F., additional, Valdizan, E., additional, Hurle, M., additional, Nistal, J., additional, Vera, V., additional, Karuppasamy, P., additional, Chaubey, S., additional, Dew, T., additional, Sherwood, R., additional, Desai, J., additional, John, L., additional, Marber, M., additional, Kunst, G., additional, Cipolletta, E., additional, Attanasio, A., additional, Del Giudice, C., additional, Campiglia, P., additional, Illario, M., additional, Berezin, A., additional, Koretskaya, E., additional, Bishop, E., additional, Fearon, I., additional, Heger, J., additional, Warga, B., additional, Abdallah, Y., additional, Meyering, B., additional, Schlueter, K., additional, Piper, H., additional, Euler, G., additional, Lavorgna, A., additional, Cecchetti, S., additional, Rio, T., additional, Coluzzi, G., additional, Carrozza, C., additional, Conti, E., additional, Andreotti, F., additional, Glavatskiy, A., additional, Uz, O., additional, Kardesoglu, E., additional, Yiginer, O., additional, Bas, S., additional, Ipcioglu, O., additional, Ozmen, N., additional, Aparci, M., additional, Cingozbay, B., additional, Ivanes, F., additional, Hillaert, M., additional, Susen, S., additional, Mouquet, F., additional, Doevendans, P., additional, Jude, B., additional, Montalescot, G., additional, Van Belle, E., additional, Castellani, C., additional, Angelini, A., additional, De Boer, O., additional, Van Der Loos, C., additional, Gerosa, G., additional, Van Der Wal, A., additional, Dumitriu, I., additional, Baruah, P., additional, Kaski, J., additional, Maytham, O., additional, D Smith, J., additional, Rose, M., additional, Cappelletti, A., additional, Pessina, A., additional, Mazzavillani, M., additional, Calori, G., additional, Margonato, A., additional, Cassese, S., additional, D'anna, C., additional, Leo, A., additional, Silenzi, A., additional, Baca', M., additional, Biasucci, L., additional, Baller, D., additional, Gleichmann, U., additional, Holzinger, J., additional, Bitter, T., additional, Horstkotte, D., additional, Antonopoulos, A., additional, Miliou, A., additional, Triantafyllou, C., additional, Masson, W., additional, Siniawski, D., additional, Sorroche, P., additional, Casanas, L., additional, Scordo, W., additional, Krauss, J., additional, Cagide, A., additional, Huang, T., additional, Wiedon, A., additional, Lee, S., additional, Walker, K., additional, O'dea, K., additional, Perez Berbel, P., additional, Arrarte Esteban, V., additional, Garcia Valentin, M., additional, Sola Villalpando, M., additional, Lopez Vaquero, C., additional, Caballero, L., additional, Quintanilla Tello, M., additional, Sogorb Garri, F., additional, Duerr, G., additional, Elhafi, N., additional, Bostani, T., additional, Swieny, L., additional, Kolobara, E., additional, Welz, A., additional, Roell, W., additional, Dewald, O., additional, Kaludercic, N., additional, Takimoto, E., additional, Nagayama, T., additional, Chen, K., additional, Shih, J., additional, Kass, D., additional, Di Lisa, F., additional, Paolocci, N., additional, Vinet, L., additional, Pezet, M., additional, Briec, F., additional, Previlon, M., additional, Rouet-Benzineb, P., additional, Hivonnait, A., additional, Charpentier, F., additional, Mercadier, J., additional, Cobo, M., additional, Llano, M., additional, Montalvo, C., additional, Exposito, V., additional, Meems, L., additional, Westenbrink, B., additional, Biesmans, L., additional, Bito, V., additional, Driessen, R., additional, Huysmans, C., additional, Mourouzis, I., additional, Pantos, C., additional, Galanopoulos, G., additional, Gavra, M., additional, Perimenis, P., additional, Spanou, D., additional, Cokkinos, D., additional, Panasenko, T., additional, Partsch, S., additional, Harjung, C., additional, Bogdanova, A., additional, Mihov, D., additional, Mocharla, P., additional, Yakushev, S., additional, Vogel, J., additional, Gassmann, M., additional, Tavakoli, R., additional, Johansen, D., additional, Sanden, E., additional, Xi, C., additional, Sundset, R., additional, Ytrehus, K., additional, Bliksoen, M., additional, Rutkovskiy, A., additional, Mariero, L., additional, Vaage, I., additional, Stenslokken, K., additional, Pisarenko, O., additional, Shulzhenko, V., additional, Studneva, I., additional, Serebryakova, L., additional, Tskitishvili, O., additional, Pelogeykina, Y., additional, Timoshin, A., additional, Vanin, A., additional, Ziberna, L., additional, Lunder, M., additional, Drevensek, G., additional, Passamonti, S., additional, Gorza, L., additional, Ravara, B., additional, Scapin, C., additional, Vitadello, M., additional, Zigrino, F., additional, Gwathmey, J., additional, Del Monte, F., additional, Vilahur, G., additional, Juan-Babot, O., additional, Onate, B., additional, Casani, L., additional, Lemoine, S., additional, Calmettes, G., additional, Jaspard-Vinassa, B., additional, Duplaa, C., additional, Couffinhal, T., additional, Diolez, P., additional, Dos Santos, P., additional, Fusco, A., additional, Sorriento, D., additional, Cervero, P., additional, Feliciello, A., additional, Barnucz, E., additional, Kozichova, K., additional, Hlavackova, M., additional, Neckar, J., additional, Kolar, F., additional, Novakova, O., additional, Novak, F., additional, Barsanti, C., additional, Abraham, N., additional, Muntean, D., additional, Mirica, S., additional, Duicu, O., additional, Raducan, A., additional, Hancu, M., additional, Fira-Mladinescu, O., additional, Ordodi, V., additional, Voelkl, J., additional, Haubner, B., additional, Neely, G., additional, Moriell, C., additional, Seidl, S., additional, Pachinger, O., additional, Penninger, J., additional, and Metzler, B., additional

Published
2010
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41. Structural and functional insights into nitrosoglutathione reductase from Chlamydomonas reinhardtii

Author

Andrea Tagliani, Jacopo Rossi, Christophe H. Marchand, Marcello De Mia, Daniele Tedesco, Libero Gurrieri, Maria Meloni, Giuseppe Falini, Paolo Trost, Stéphane D. Lemaire, Simona Fermani, and Mirko Zaffagnini

Subjects
Chlamydomonas, Cysteine, Nitrosoglutathione, Nitrosoglutathione reductase, Redox regulation, Thiol modification, Medicine (General), R5-920, Biology (General), QH301-705.5
Abstract

Protein S-nitrosylation plays a fundamental role in cell signaling and nitrosoglutathione (GSNO) is considered as the main nitrosylating signaling molecule. Enzymatic systems controlling GSNO homeostasis are thus crucial to indirectly control the formation of protein S-nitrosothiols. GSNO reductase (GSNOR) is the key enzyme controlling GSNO levels by catalyzing its degradation in the presence of NADH. Here, we found that protein extracts from the microalga Chlamydomonas reinhardtii catabolize GSNO via two enzymatic systems having specific reliance on NADPH or NADH and different biochemical features. Scoring the Chlamydomonas genome for orthologs of known plant GSNORs, we found two genes encoding for putative and almost identical GSNOR isoenzymes. One of the two, here named CrGSNOR1, was heterologously expressed and purified. Its kinetic properties were determined and the three-dimensional structures of the apo-, NAD+- and NAD+/GSNO-forms were solved. These analyses revealed that CrGSNOR1 has a strict specificity towards GSNO and NADH, and a conserved folding with respect to other plant GSNORs. The catalytic zinc ion, however, showed an unexpected variability of the coordination environment. Furthermore, we evaluated the catalytic response of CrGSNOR1 to thermal denaturation, thiol-modifying agents and oxidative modifications as well as the reactivity and position of accessible cysteines. Despite being a cysteine-rich protein, CrGSNOR1 contains only two solvent-exposed/reactive cysteines. Oxidizing and nitrosylating treatments have null or limited effects on CrGSNOR1 activity and folding, highlighting a certain resistance of the algal enzyme to redox modifications. The molecular mechanisms and structural features underlying the response to thiol-based modifications are discussed.

Published
2021
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48. The Thioredoxin-Regulated α-Amylase 3 of Arabidopsis thaliana Is a Target of S-Glutathionylation

Author

Libero Gurrieri, Luca Distefano, Claudia Pirone, Daniel Horrer, David Seung, Mirko Zaffagnini, Nicolas Rouhier, Paolo Trost, Diana Santelia, and Francesca Sparla

Subjects
α-amylase 3, post-translational redox modifications, S-glutathionylation, disulfide, cysteine pKa, glutaredoxin, Plant culture, SB1-1110
Abstract

Reactive oxygen species (ROS) are produced in cells as normal cellular metabolic by-products. ROS concentration is normally low, but it increases under stress conditions. To stand ROS exposure, organisms evolved series of responsive mechanisms. One such mechanism is protein S-glutathionylation. S-glutathionylation is a post-translational modification typically occurring in response to oxidative stress, in which a glutathione reacts with cysteinyl residues, protecting them from overoxidation. α-Amylases are glucan hydrolases that cleave α-1,4-glucosidic bonds in starch. The Arabidopsis genome contains three genes encoding α-amylases. The sole chloroplastic member, AtAMY3, is involved in osmotic stress response and stomatal opening and is redox-regulated by thioredoxins. Here we show that AtAMY3 activity was sensitive to ROS, such as H2O2. Treatments with H2O2 inhibited enzyme activity and part of the inhibition was irreversible. However, in the presence of glutathione this irreversible inhibition was prevented through S-glutathionylation. The activity of oxidized AtAMY3 was completely restored by simultaneous reduction by both glutaredoxin (specific for the removal of glutathione-mixed disulfide) and thioredoxin (specific for the reduction of protein disulfide), supporting a possible liaison between both redox modifications. By comparing free cysteine residues between reduced and GSSG-treated AtAMY3 and performing oxidation experiments of Cys-to-Ser variants of AtAMY3 using biotin-conjugated GSSG, we could demonstrate that at least three distinct cysteinyl residues can be oxidized/glutathionylated, among those the two previously identified catalytic cysteines, Cys499 and Cys587. Measuring the pKa values of the catalytic cysteines by alkylation at different pHs and enzyme activity measurement (pKa1 = 5.70 ± 0.28; pKa2 = 7.83 ± 0.12) showed the tendency of one of the two catalytic cysteines to deprotonation, even at physiological pHs, supporting its propensity to undergo redox post-translational modifications. Taking into account previous and present findings, a functional model for redox regulation of AtAMY3 is proposed.

Published
2019
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49. Impact of Drought on Soluble Sugars and Free Proline Content in Selected Arabidopsis Mutants

Author

Libero Gurrieri, Martina Merico, Paolo Trost, Giuseppe Forlani, and Francesca Sparla

Subjects
drought stress, metabolic adjustment, proline, soluble sugars, water-insoluble carbohydrates, Biology (General), QH301-705.5
Abstract

Water shortage is an increasing problem affecting crop yield. Accumulation of compatible osmolytes is a typical plant response to overcome water stress. Sucrose synthase 1 (SUS1), and glucan, water dikinase 2 (GWD2) and δ1-pyrroline-5-carboxylate synthetase 1 (P5CS1) are members of small protein families whose role in the response of Arabidopsis thaliana plants to mild osmotic stress has been studied in this work. Comparative analysis between wild-type and single loss-of-function T-DNA plants at increasing times following exposure to drought showed no differences in the content of water-insoluble carbohydrate (i.e., transitory starch and cell wall carbohydrates) and in the total amount of amino acids. On the contrary, water-soluble sugars and proline contents were significantly reduced compared to wild-type plants regardless of the metabolic pathway affected by the mutation. The present results contribute to assigning a physiological role to GWD2, the least studied member of the GWD family; strengthening the involvement of SUS1 in the response to osmotic stress; showing a greater contribution of soluble sugars than proline in osmotic adjustment of Arabidopsis in response to drought. Finally, an interaction between proline and soluble sugars emerged, albeit its nature remains speculative and further investigations will be required for complete comprehension.

Published
2020
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