Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Maldonado‐Cervantes, Enrique; Huerta‐Ocampo, José Ángel; Montero-Morán, Gabriela M.; Barrera-Pacheco, Alberto; Espitia-Rangel, Eduardo; Rosa, Ana

doi:10.1016/j.jcs.2014.02.008

Cited by 17 publications

(7 citation statements)

References 27 publications

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“…AcLEA cDNA contains an ORF of 516 bp that codifies for a 172 amino acids protein with a molecular mass calculated of 18.34 kDa and a theoretical p I of 8.58, values that corresponded to experimental data previously reported (Maldonado-Cervantes et al, 2014). The sequence (Supplementary Figure S2A) was deposited in the GenBank with access code KX852451.…”

Section: Resultssupporting

confidence: 85%

“…Bioinformatics analyses, using LC-MS/MS information (Maldonado-Cervantes et al, 2014) and the A. hypochondriacus transcriptome (Delano-Frier et al, 2011), allowed us to design specific primers for cloning the full-length AcLEA cDNA. Amplified AcLEA fragment was ligated into pET28mod vector (Supplementary Figures S1A,B).…”

Section: Resultsmentioning

confidence: 99%

“…Immature seeds (15 days after anthesis) of Amaranthus cruentus were used to extract total RNA with TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) and cDNA was synthesized as previously reported (Maldonado-Cervantes et al, 2014). AcLEA cDNA was amplified using specific primers containing Nde I (5′-CATATGGCATCACATGGTCAGAGT-3′) and Xho I (5′-CTCGAGCTAGGGCCTAGTAGTCTTAATTGGATC-3′) restriction sites.…”

Section: Methodsmentioning

confidence: 99%

“…Amaranth is resistant against several types of stresses such as pest (Valdes-Rodríguez et al, 2007), heat (Maughan et al, 2009), drought (Huerta-Ocampo et al, 2011), and salinity (Aguilar-Hernández et al, 2011; Huerta-Ocampo et al, 2014). The recent report of Amaranthus cruentus seed proteome by 2D-PAGE revealed the over-accumulation of one spot identified as a LEA protein (Maldonado-Cervantes et al, 2014). In the present study, we have cloned the corresponding LEA cDNA from A. cruentus ( AcLEA , GenBank accession no.…”

Section: Introductionmentioning

confidence: 99%

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Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

et al. 2017

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Late embryogenesis abundant (LEA) proteins are part of a large protein family that protect other proteins from aggregation due to desiccation or osmotic stresses. Recently, the Amaranthus cruentus seed proteome was characterized by 2D-PAGE and one highly accumulated protein spot was identified as a LEA protein and was named AcLEA. In this work, AcLEA cDNA was cloned into an expression vector and the recombinant protein was purified and characterized. AcLEA encodes a 172 amino acid polypeptide with a predicted molecular mass of 18.34 kDa and estimated pI of 8.58. Phylogenetic analysis revealed that AcLEA is evolutionarily close to the LEA3 group. Structural characteristics were revealed by nuclear magnetic resonance and circular dichroism methods. We have shown that recombinant AcLEA is an intrinsically disordered protein in solution even at high salinity and osmotic pressures, but it has a strong tendency to take a secondary structure, mainly folded as α-helix, when an inductive additive is present. Recombinant AcLEA function was evaluated using Escherichia coli as in vivo model showing the important protection role against desiccation, oxidant conditions, and osmotic stress. AcLEA recombinant protein was localized in cytoplasm of Nicotiana benthamiana protoplasts and orthologs were detected in seeds of wild and domesticated amaranth species. Interestingly AcLEA was detected in leaves, stems, and roots but only in plants subjected to salt stress. This fact could indicate the important role of AcLEA protection during plant stress in all amaranth species studied.

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Section: Resultssupporting

confidence: 85%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

et al. 2017

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“…Proteins from the six conditions (50 μg) were resolved in a short migration of about 4 mm in a 13.5% SDS-PAGE and stained with Coomassie blue. Protein bands were excised from the gel, distained, reduced with DTT followed by protein alkylation with iodoacetamide and finally digested overnight with trypsin as previously reported by Maldonado-Cervantes et al (2014). Next, peptide samples were desalted with C18 cartridges (Sep-Pak C18, Waters, Milford, MA, USA) and dried under vacuum.…”

Section: Identification Of Proteins By Shotgun-type Protein Analysismentioning

confidence: 99%

Interaction of the human intestinal microbiota with the release of bound phenolic compounds in chickpea (Cicer arietinum L.)

Perez‐Perez

Huerta‐Ocampo

Ramos-Enríquez

et al. 2021

Int J of Food Sci Tech

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The objective of this work was to release bound phenolic compounds (PC) from chickpea by the interaction of the microbiota of a volunteer and to identify the enzymes implied to deliver these PC. The highest amount of PC was released at 12 and 24 h (16.8-18.5 mg GAE/g). Higher antioxidant capacity was observed in these hours through 2,20-azino-bis-(3ethylbenzothiazoline-6-sulfonic acid), 2,2-diphenyl-1picrylhydrazyl, ferric reducing antioxidant power and 2,´2.Azobis (2 methylpropionamidine) techniques. Escherichia, Klebsiella, Bacteroides and Veillonella were some genera identified in the microbiota implied in delivered PC. The principal PC identified by ultra performance liquid chromatography-mass spectrometry were flavonoids. Proteomic analysis identified hundreds of proteins from the intestinal microbiota after 12 h of fermentation, including enzymes related to the release of bound PC from chickpea such as pectin esterase. Therefore, this enzyme could be used in other food sources for the release of PC bound to the food matrix and thus take advantage of their bioactive benefits.

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Proteins and Amino Acids of Kernels

et al. 2017

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Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Cited by 17 publications

References 27 publications

Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

Interaction of the human intestinal microbiota with the release of bound phenolic compounds in chickpea (Cicer arietinum L.)

Proteins and Amino Acids of Kernels

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