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1.
Toxicol In Vitro ; 50: 347-372, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29626626

RESUMEN

Silver nanoparticles (AgNPs) have been incorporated into several consumer products. While these advances in technology are promising and exciting, the effects of these nanoparticles have not equally been studied. Due to the size, AgNPs can penetrate the body through oral exposure and reach the gastrointestinal tract. The present study was designed as a comparative proteomic analysis of Caco-2 cells, used as an in vitro model of the small intestine, exposed to 30 nm citrate stabilized-silver nanoparticles (AgNPs) for 24 or 72 h. Using two complementary proteomic approaches, 2D gel-based and label-free mass spectrometry, we present insight into the effects of AgNPs at proteins level. Exposure of 1 or 10 µg/mL AgNPs to Caco-2 cells resulted in 56 and 88 altered proteins at 24 h and 72 h respectively, by 2D gel-based technique. Ten of these proteins were found to be common between the two time-points. Using label-free mass spectrometry technique, 291 and 179 altered proteins were found at 24 h and 72 h, of which 24 were in common. Analysis of the proteomes showed several major biological processes altered, from which, cell cycle, cell morphology, cellular function and maintenance were the most affected.


Asunto(s)
Nanopartículas del Metal/toxicidad , Proteoma/efectos de los fármacos , Plata/toxicidad , Células CACO-2 , Supervivencia Celular/efectos de los fármacos , Humanos , Intestino Delgado/metabolismo , Proteómica , Nitrato de Plata/toxicidad
2.
DNA Repair (Amst) ; 50: 14-21, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-28017527

RESUMEN

This pilot study was carried out to assess the effect of radio-contaminated Chernobyl environment on plant genome integrity 27 years after the accident. For this purpose, nuclei were isolated from root tips of the soybean seedlings harvested from plants grown in the Chernobyl area for seven generations. Neutral, neutral-alkaline, and methylation-sensitive comet assays were performed to evaluate the induction and repair of primary DNA damage and the epigenetic contribution to stress adaptation mechanisms. An increased level of single and double strand breaks in the radio-contaminated Chernobyl seedlings at the stage of primary root development was detected in comparison to the controls. However, the kinetics of the recovery of DNA breaks of radio-contaminated Chernobyl samples revealed that lesions were efficiently repaired at the stage of cotyledon. Methylation-sensitive comet assay revealed comparable levels in the CCGG methylation pattern between control and radio-contaminated samples with a slight increase of approximately 10% in the latter ones. The obtained preliminary data allow us to speculate about the onset of mechanisms providing an adaptation potential to the accumulated internal irradiation after the Chernobyl accident. Despite the limitations of this study, we showed that comet assay is a sensitive and flexible technique which can be efficiently used for genotoxic screening of plant specimens in natural and human-made radio-contaminated areas, as well as for safety monitoring of agricultural products.


Asunto(s)
Adaptación Fisiológica/genética , Metilación de ADN , Reparación del ADN , ADN de Plantas/metabolismo , Epigénesis Genética , Glycine max/genética , Accidente Nuclear de Chernóbil , Ensayo Cometa , Roturas del ADN de Doble Cadena , Roturas del ADN de Cadena Simple , Cinética , Proyectos Piloto , Plantones/genética , Plantones/metabolismo , Plantones/fisiología , Glycine max/metabolismo , Glycine max/fisiología
3.
Front Plant Sci ; 6: 493, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26217350

RESUMEN

Plants have the ability to grow and successfully reproduce in radio-contaminated environments, which has been highlighted by nuclear accidents at Chernobyl (1986) and Fukushima (2011). The main aim of this article is to summarize the advances of the Chernobyl seed project which has the purpose to provide proteomic characterization of plants grown in the Chernobyl area. We present a summary of comparative proteomic studies on soybean and flax seeds harvested from radio-contaminated Chernobyl areas during two successive generations. Using experimental design developed for radio-contaminated areas, altered abundances of glycine betaine, seed storage proteins, and proteins associated with carbon assimilation into fatty acids were detected. Similar studies in Fukushima radio-contaminated areas might complement these data. The results from these Chernobyl experiments can be viewed in a user-friendly format at a dedicated web-based database freely available at http://www.chernobylproteomics.sav.sk.

4.
Front Plant Sci ; 6: 433, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26124766

RESUMEN

The aim of the work was to test a relatively simple proteomics approach based on phenol extraction and two-dimensional gel electrophoresis (2-DE) with 7 cm immobilized pH gradient strips for the determination of clinically relevant proteins in wheat grain. Using this approach, 157 2-DE spots were quantified in biological triplicate, out of which 55 were identified by matrix-assisted laser desorption/ionization - time of flight tandem mass spectrometry. Clinically relevant proteins associated with celiac disease, wheat dependent exercise induced anaphylaxis, baker's asthma, and food allergy, were detected in 24 2-DE spots. However, alcohol-soluble gliadins were not detected with this approach. The comparison with a recent quantitative study suggested that gel-based and gel-free proteomics approaches are complementary for the detection and quantification of clinically relevant proteins in wheat grain.

6.
Proteomics ; 15(10): 1631-7, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25865070

RESUMEN

The International Plant Proteomics Organization (INPPO) is a global platform of the plant proteomics community or, more generally, the scientific community that uses proteomics to address plant biology. Organizing an international conference is one of its initiatives to promote plant proteomics by involving and gathering scientists/researchers/students and by disseminating the acquired knowledge. In this fourth INPPO Highlights, the first INPPO World Congress 2014 (INPPO2014) is described and discussed. The INPPO2014 was held at the University of Hamburg (Germany) with the title "Plant Proteomics: Methodology to Biology" under the leadership of Sabine Lüthje (Germany). Participants (around 150) from 38 nations attended this congress covering all continents. The four-day scientific program comprised 52 lectures and 61 poster presentations in a highly professional and friendly atmosphere on mass spectrometry and gel-based proteomics. Two round-table open discussions deliberated on plant proteomics, its associated international organizations/initiatives and future INPPO perspectives. The Second INPPO World Congress 2016 (INPPO2016) "The Quest for Tolerant Varieties-Phenotyping at Plant and Cellular Level" is planned to be organized in Bratislava (Slovakia) under the leadership of Martin Hajduch (Slovak Republic) and Sébastien Carpentier (Belgium) and cosponsored by the COST action FA1306.


Asunto(s)
Conocimiento , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Proteómica , Apoyo a la Investigación como Asunto/economía
7.
Front Plant Sci ; 6: 1215, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26793203

RESUMEN

Plants continue to flourish around the site of the Chernobyl Nuclear Power Plant disaster. The ability of plants to transcend the radio-contaminated environment was not anticipated and is not well understood. The aim of this study was to evaluate the proteome of flax (Linum usitatissimum L.) during seed filling by plants grown for a third generation near Chernobyl. For this purpose, seeds were harvested at 2, 4, and 6 weeks after flowering and at maturity, from plants grown in either non-radioactive or radio-contaminated experimental fields. Total proteins were extracted and the two-dimensional gel electrophoresis (2-DE) patterns analyzed. This approach established paired abundance profiles for 130 2-DE spots, e.g., profiles for the same spot across seed filling in non-radioactive and radio-contaminated experimental fields. Based on Analysis of Variance (ANOVA) followed by sequential Bonferroni correction, eight of the paired abundance profiles were discordant. Results from tandem mass spectrometry show that four 2-DE spots are discordant because they contain fragments of the cupin superfamily-proteins. Most of the fragments were derived from the N-terminal half of native cupins. Revisiting previously published data, it was found that cupin-fragments were also involved with discordance in paired abundance profiles of second generation flax seeds. Based on these observations we present an updated working model for the growth and reproductive success of flax in a radio-contaminated Chernobyl environment. This model suggests that the increased abundance of cupin fragments or isoforms and monomers contributes to the successful growth and reproduction of flax in a radio-contaminated environment.

8.
J Agric Food Chem ; 62(47): 11547-56, 2014 Nov 26.
Artículo en Inglés | MEDLINE | ID: mdl-25365400

RESUMEN

Although ginkgo (Maidenhair tree, Ginkgo biloba L.) is an ancient medicinal and ornamental tree, there has not previously been any systematic proteomic study of the leaves. Herein we describe results from the initial study identifying abundant ginkgo leaf proteins and present a gel reference map. Proteins were isolated from fully developed mature leaves in biological triplicate and analyzed by two-dimensional electrophoresis plus tandem mass spectrometry. Using this approach, we were able to reproducibly quantify 190 abundant protein spots, from which 157 proteins were identified. Most of identified proteins are associated with the energy and protein destination/storage categories. The reference map provides a basis for understanding the accumulation of flavonoids and other phenolic compounds in mature leaves (e.g., identification of chalcone synthase, the first committed enzyme in flavonoid biosynthesis). We additionally detected several proteins of as yet unknown function. These proteins comprise a pool of potential targets that might be useful in nontraditional medical applications.


Asunto(s)
Ginkgo biloba/química , Hojas de la Planta/química , Proteoma/metabolismo , Cromatografía Liquida , Electroforesis en Gel Bidimensional , Flavonoides/análisis , Fenoles/análisis , Proteínas de Plantas/química , Proteómica , Espectrometría de Masas en Tándem
9.
J Proteomics ; 104: 57-65, 2014 Jun 02.
Artículo en Inglés | MEDLINE | ID: mdl-24530378

RESUMEN

Totipotency, the ability of somatic plant cell to generate whole plant through somatic embryogenesis, is still not well understood. In this study, maize immature zygotic embryos were used to generate embryogenic (EC) and non-embryogenic (NEC) calli. In order to compare proteomes of EC and NEC, two-dimensional electrophoresis (2-DE) in combination with mass spectrometry was used. This approach resulted into 361 quantified 2-DE spots out of which 44 were found statistically significantly differentially abundant between EC and NEC. Mass spectrometry provided the identity for 23 proteins that were classified into 8 metabolic categories. The most abundant were proteins associated with energy followed by proteins associated with disease and defense. Based on the abundances of identified proteins in this and other studies, working model for plant totipotency was proposed. One aspect of this working model suggests that increased abundances of proteins associated with pyruvate biosynthesis and suppression of embryogenic genes might be responsible for differences between EC and NEC cells. Furthermore we speculate that the increased abundance of lipoxygenase in the NEC cells results in changes in the equilibrium levels of one or more signaling molecules and is at least partly responsible for somatic cell reprogramming during totipotency. BIOLOGICAL SIGNIFICANCE: Totipotency, the ability of somatic plant cell to generate whole plant through somatic embryogenesis, is still not well understood. In order to further advance understanding of this biological phenomenon, proteomes of embryogenic and non-embryogenic callus, derived from immature zygotic embryos of inbred maize line A19, were compared using 2-DE based proteomic technology. Based on the abundances of identified proteins in this and other studies, working model for plant totipotency was proposed. One aspect of this working model suggests that increased abundances of proteins associated with pyruvate biosynthesis and suppression of embryogenic genes might be responsible for differences between EC and NEC cells. Furthermore we speculate that the increased abundance of lipoxygenase in the NEC cells results in changes in the equilibrium levels of one or more signaling molecules and is at least partly responsible for somatic cell reprogramming during totipotency. This article is part of a Special Issue entitled: Environmental and structural proteomics.


Asunto(s)
Desarrollo Embrionario/fisiología , Células Madre Embrionarias/metabolismo , Oxilipinas/metabolismo , Semillas/metabolismo , Células Madre Totipotentes/metabolismo , Zea mays/embriología , Zea mays/metabolismo , Células Madre Embrionarias/citología , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Semillas/citología , Semillas/crecimiento & desarrollo , Células Madre Totipotentes/citología , Zea mays/citología
10.
Methods Mol Biol ; 1072: 555-61, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24136546

RESUMEN

Two serious nuclear accidents during the past quarter of a century contaminated large agricultural areas with radioactivity. The remediation and possible recovery of radio-contaminated areas for agricultural purposes require comprehensive characterization of plants grown in such places. Here we describe the quantitative proteomics method that we use to analyze proteins isolated from seeds of plants grown in radioactive Chernobyl zone.


Asunto(s)
Accidente Nuclear de Chernóbil , Proteómica/métodos , Contaminantes Radiactivos del Suelo/química , Electroforesis en Gel de Poliacrilamida , Focalización Isoeléctrica , Espectrometría de Masas , Péptido Hidrolasas/metabolismo , Fenoles , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Glycine max/metabolismo
11.
J Proteome Res ; 12(11): 4799-806, 2013 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-24111740

RESUMEN

Starting in 2007, we have grown soybean (Glycine max [L.] Merr. variety Soniachna) and flax (Linum usitatissimum, L. variety Kyivskyi) in the radio-contaminated Chernobyl area and analyzed the seed proteomes. In the second-generation flax seeds, we detected a 12% increase in oil content. To characterize the bases for this increase, seed development has been studied. Flax seeds were harvested in biological triplicate at 2, 4, and 6 weeks after flowering and at maturity from plants grown in nonradioactive and radio-contaminated plots in the Chernobyl area for two generations. Quantitative proteomic analyses based on 2-D gel electrophoresis (2-DE) allowed us to establish developmental profiles for 199 2-DE spots in both plots, out of which 79 were reliably identified by tandem mass spectrometry. The data suggest a statistically significant increased abundance of proteins associated with pyruvate biosynthesis via cytoplasmic glycolysis, L-malate decarboxylation, isocitrate dehydrogenation, and ethanol oxidation to acetaldehyde in early stages of seed development. This was followed by statistically significant increased abundance of ketoacyl-[acylcarrier protein] synthase I related to condensation of malonyl-ACP with elongating fatty acid chains. On the basis of these and previous data, we propose a preliminary model for plant adaptation to growth in a radio-contaminated environment. One aspect of the model suggests that changes in carbon assimilation and fatty acid biosynthesis are an integral part of plant adaptation.


Asunto(s)
Adaptación Biológica/efectos de la radiación , Accidente Nuclear de Chernóbil , Lino/genética , Lino/efectos de la radiación , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Modelos Biológicos , Proteoma/efectos de la radiación , Adaptación Biológica/genética , Carbono/metabolismo , Radioisótopos de Cesio/análisis , Electroforesis en Gel Bidimensional , Lino/química , Regulación de la Expresión Génica de las Plantas/genética , Glucólisis , Aceite de Linaza/análisis , Proteoma/genética , Ácido Pirúvico/metabolismo , Semillas/química , Semillas/crecimiento & desarrollo , Semillas/efectos de la radiación , Suelo/química , Radioisótopos de Estroncio/análisis , Espectrometría de Masas en Tándem , Ucrania
12.
J Proteome Res ; 12(11): 4862-9, 2013 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-24007624

RESUMEN

The amount of clinically relevant, allergy-related proteins in wheat grain is still largely unknown. The application of proteomics may create a platform not only for identification and characterization, but also for quantitation of these proteins. The aim of this study was to evaluate the data-independent quantitative mass spectrometry (MS(E)) approach in combination with 76 wheat allergenic sequences downloaded from the AllergenOnline database ( www.allergenonline.org ) as a starting point. Alcohol soluble extracts of gliadin and glutenin proteins were analyzed. This approach has resulted in identification and quantification of 15 allergenic protein isoforms that belong to amylase/trypsin inhibitors, γ-gliadins, and high or low molecular weight glutenins. Additionally, several peptides carrying four previously discovered epitopes of γ-gliadin B precursor have been detected. These data were validated against the UniProt database, which contained 11764 Triticeae protein sequences. The identified allergens are discussed in relation to Baker's asthma, food allergy, wheat dependent exercise induced anaphylaxis, atopic dermatitis, and celiac disease (i.e., gluten-sensitive enteropathy). In summary, the results showed that the MS(E) approach is suitable for quantitative analysis and allergens profiling in wheat varieties and/or other food matrices.


Asunto(s)
Epítopos/genética , Extractos Vegetales/genética , Proteínas de Plantas/metabolismo , Triticum/química , Antígenos de Plantas/genética , Antígenos de Plantas/metabolismo , Bases de Datos Genéticas , Espectrometría de Masas/métodos , Proteínas de Plantas/genética , Proteómica
13.
J Proteomics ; 93: 65-73, 2013 Nov 20.
Artículo en Inglés | MEDLINE | ID: mdl-23268118

RESUMEN

Precise content of gliadin (Glia) and glutenin (Glu) proteins in wheat grain are largely unknown despite their association with celiac disease, various allergies, and physical processing properties of wheat. Developing methods to quantitatively measure clinically relevant proteins could support advancement in understanding exposure thresholds and clinical study design. The aim of this study was to use a data-independent mass spectrometry (MS(E)) approach for quantifying gliadin and glutenin proteins in wheat grain. The biologically replicated analysis yielded concentrations for 34 gliadin and 22 glutenin proteins. The primary focus of this survey was on measuring celiac disease proteins and baker's asthma associated proteins along with the proteins associated with viscoelastic properties of wheat flour and grain texture. The technical coefficients of variation ranged from 0.12 to 1.39 and indicate that MS(E) proteomics is a reproducible quantitative method for the determination of gliadin and glutenin content in the highly complex matrix of protein extracts from wheat grain. This article is part of a Special Issue entitled: Translational Plant Proteomics.


Asunto(s)
Asma/genética , Enfermedad Celíaca/genética , Gliadina/análisis , Glútenes/análisis , Espectrometría de Masas/métodos , Triticum/química , Proteómica
14.
PLoS One ; 7(10): e48169, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23110204

RESUMEN

Plants grow and reproduce in the radioactive Chernobyl area, however there has been no comprehensive characterization of these activities. Herein we report that life in this radioactive environment has led to alteration of the developing soybean seed proteome in a specific way that resulted in the production of fertile seeds with low levels of oil and ß-conglycinin seed storage proteins. Soybean seeds were harvested at four, five, and six weeks after flowering, and at maturity from plants grown in either non-radioactive or radioactive plots in the Chernobyl area. The abundance of 211 proteins was determined. The results confirmed previous data indicating that alterations in the proteome include adaptation to heavy metal stress and mobilization of seed storage proteins. The results also suggest that there have been adjustments to carbon metabolism in the cytoplasm and plastids, increased activity of the tricarboxylic acid cycle, and decreased condensation of malonyl-acyl carrier protein during fatty acid biosynthesis.


Asunto(s)
Carbono/metabolismo , Accidente Nuclear de Chernóbil , Glycine max/metabolismo , Metales Pesados/toxicidad , Semillas/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/genética , Proteínas de Plantas/metabolismo , Plastidios/genética , Proteoma/efectos de los fármacos , Proteoma/metabolismo , Proteínas de Almacenamiento de Semillas/metabolismo , Semillas/efectos de los fármacos , Glycine max/efectos de los fármacos
15.
Front Plant Sci ; 3: 231, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23087698

RESUMEN

Two serious nuclear accidents during the last quarter century (Chernobyl, 1986 and Fukushima, 2011) contaminated large agricultural areas with radioactivity. The database "Seeds in Chernobyl" (http://www.chernobylproteomics.sav.sk) contains the information about the abundances of hundreds of proteins from on-going investigation of mature and developing seed harvested from plants grown in radioactive Chernobyl area. This database provides a useful source of information concerning the response of the seed proteome to permanently increased level of ionizing radiation in a user-friendly format.

16.
J Proteome Res ; 11(1): 372-85, 2012 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-22136409

RESUMEN

Flooding injury is a major problem in soybean cultivation. A proteomics approach was used to clarify the occurrence of changes in protein expression level and phosphorylation in soybeans under flooding stress. Two-day-old seedlings were flooded for 1 day, proteins were extracted from root tips of the seedlings and digested with trypsin, and their expression levels and phosphorylation states were compared to those of untreated controls using mass spectrometry-based proteomics techniques. Phosphoproteins were enriched using a phosphoprotein purification column prior to digestion and mass spectrometry. The expression of proteins involved in energy production increased as a result of flooding, while expression of proteins involved in protein folding and cell structure maintenance decreased. Flooding induced changes of phosphorylation status of proteins involved in energy generation, protein synthesis and cell structure maintenance. The response to flooding stress may be regulated by both modulation of protein expression and phosphorylation state. Energy-demanding and production-related metabolic pathways may be particularly subject to regulation by changes in protein phosphorylation during flooding.


Asunto(s)
Glycine max/fisiología , Meristema/fisiología , Fosfoproteínas/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Estrés Fisiológico , Inundaciones , Regulación de la Expresión Génica de las Plantas , Meristema/enzimología , Meristema/metabolismo , Redes y Vías Metabólicas/genética , Fragmentos de Péptidos/química , Mapeo Peptídico , Fosfoproteínas/química , Fosfoproteínas/genética , Proteínas de Plantas/genética , Proteoma/química , Proteoma/genética , Proteómica , Piruvato Quinasa/química , Piruvato Quinasa/genética , Piruvato Quinasa/metabolismo , Glycine max/enzimología , Glycine max/metabolismo , Espectrometría de Masas en Tándem , Transcripción Genética
17.
J Proteomics ; 74(10): 1974-84, 2011 Sep 06.
Artículo en Inglés | MEDLINE | ID: mdl-21616182

RESUMEN

Coxiella burnetii, a category B biological warfare agent, causes multiple outbreaks of the zoonotic disease Q fever world-wide, each year. The virulent phase I and avirulent phase II variants of the Nine Mile RSA 493 and 439 strains of C. burnetii were propagated in embryonated hen eggs and then purified by centrifugation through Renografin gradients. Total protein fractions were isolated from each phase and subjected to analysis by one-dimensional electrophoresis plus tandem mass spectrometry. A total of 235 and 215 non-redundant proteins were unambiguously identified from the phase I and II cells, respectively. Many of these proteins had not been previously reported in proteomic studies of C. burnetii. The newly identified proteins should provide additional insight into the pathogenesis of Q fever. Several of the identified proteins are involved in the biosynthesis and metabolism of components of the extracellular matrix. Forty-four of the proteins have been annotated as having distinct roles in the pathogenesis or survival of C. burnetii within the harsh phagolysosomal environment. We propose that nine enzymes specifically involved with lipopolysaccharide biosynthesis and metabolism, and that are distinctively present in phase I cells, are virulence-associated proteins.


Asunto(s)
Proteínas Bacterianas/análisis , Coxiella burnetii/patogenicidad , Fiebre Q/inmunología , Antígenos Bacterianos/genética , Antígenos Bacterianos/aislamiento & purificación , Proteínas Bacterianas/aislamiento & purificación , Coxiella burnetii/genética , Lipopolisacáridos/biosíntesis , Lipopolisacáridos/metabolismo , Proteómica , Fiebre Q/etiología , Espectrometría de Masas en Tándem , Virulencia/genética
18.
J Proteomics ; 74(8): 1378-84, 2011 Aug 12.
Artículo en Inglés | MEDLINE | ID: mdl-21385628

RESUMEN

Molecular characterization of crop plants grown in remediated, formerly radioactive, areas could establish a framework for future agricultural use of these areas. Recently, we have established a quantitative reference map for mature flax seed proteins (Linum usitatissimum L.) harvested from a remediated plot in Chernobyl town. Herein we describe results from our ongoing studies of this subject, and provide a proteomics-based characterization of developing flax seeds harvested from same field. A quantitative approach, based on 2-dimensional electrophoresis (2-DE) and tandem mass spectrometry, yielded expression profiles for 379 2-DE spots through seed development. Despite the paucity of genomic resources for flax, the identity for 102 proteins was reliably determined. These proteins were sorted into 11 metabolic functional classes. Proteins of unknown function comprise the largest group, and displayed a pattern of decreased abundance throughout seed development. Analysis of the composite expression profiles for metabolic protein classes revealed specific expression patterns during seed development. For example, there was an overall decrease in abundance of the glycolytic enzymes during seed development.


Asunto(s)
Accidente Nuclear de Chernóbil , Lino/efectos de la radiación , Semillas/crecimiento & desarrollo , Agricultura , Lino/crecimiento & desarrollo , Lino/metabolismo , Perfilación de la Expresión Génica , Proteómica
19.
Proteomics ; 11(9): 1619-29, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21413150

RESUMEN

Proteomics is increasingly being used to understand enzyme expression and regulatory mechanisms involved in the accumulation of storage reserves in crops with sequenced genomes. During the past six years, considerable progress has been made to characterize proteomes of both mature and developing seeds, particularly oilseeds - plants which accumulate principally oil and protein as storage reserves. This review summarizes the emerging proteomics data, with emphasis on seed filling in soy, rapeseed, castor and Arabidopsis as each of these oilseeds were analyzed using very similar proteomic strategies. These parallel studies provide a comprehensive view of source-sink relationships, specifically sucrose assimilation into organic acid intermediates for de novo amino acid and fatty acid synthesis. We map these biochemical processes for seed maturation and illustrate the differences and similarities among the four oilseeds. For example, while the four oilseeds appear capable of producing cytosolic phosphoenolpyruvate as the principal carbon intermediate, soybean and castor also express malic enzymes and malate dehydrogenase, together capable of producing malate that has been previously proposed to be the major intermediate for fatty acid synthesis in castor. We discuss these and other differences in the context of intermediary metabolism for developing oilseeds.


Asunto(s)
Aceites de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Proteómica/métodos , Semillas/metabolismo , Metabolómica/métodos , Modelos Biológicos , Plantas/embriología , Plantas/metabolismo , Proteómica/tendencias , Semillas/embriología , Especificidad de la Especie , Factores de Tiempo
20.
Phytochemistry ; 72(10): 1308-15, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-21144539

RESUMEN

In recent years there has been an increasing tendency toward remediation of contaminated areas for agriculture purposes. The study described herein is part of a comprehensive, long-term characterization of crop plants grown in the area formerly contaminated with radioactivity. As a first step, we have established a quantitative map of proteins isolated from mature flax (Linum usitatissimum L.) seeds harvested from plants grown in a remediated plot localized directly in Chernobyl town. Flax was selected because it is a crop of economic and historical importance, despite the relative paucity of molecular resources. We used 2-dimensional electrophoresis followed by tandem mass spectrometry to establish a high-resolution seed proteome map. This approach yielded quantitative information for 318 protein spots. Genomic sequence resources for flax are very limited, leaving us with an "unknown function" annotation for 38% of the proteins analyzed including several that comprise very large spots. In addition to the seed storage proteins, we were able to reliably identify 82 proteins many of which are involved with central metabolism.


Asunto(s)
Accidente Nuclear de Chernóbil , Productos Agrícolas/metabolismo , Lino/metabolismo , Proteínas de Plantas/análisis , Semillas/crecimiento & desarrollo , Semillas/metabolismo , Productos Agrícolas/crecimiento & desarrollo , Lino/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Electroforesis Bidimensional Diferencial en Gel
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