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Ripening of pepper (Capsicum annuum) fruit is characterized by an enhancement of protein tyrosine nitration.
Ann Bot. 2015 Sep; 116(4):637-47.AB

Abstract

BACKGROUND AND AIMS

Pepper (Capsicum annuum, Solanaceae) fruits are consumed worldwide and are of great economic importance. In most species ripening is characterized by important visual and metabolic changes, the latter including emission of volatile organic compounds associated with respiration, destruction of chlorophylls, synthesis of new pigments (red/yellow carotenoids plus xanthophylls and anthocyanins), formation of pectins and protein synthesis. The involvement of nitric oxide (NO) in fruit ripening has been established, but more work is needed to detail the metabolic networks involving NO and other reactive nitrogen species (RNS) in the process. It has been reported that RNS can mediate post-translational modifications of proteins, which can modulate physiological processes through mechanisms of cellular signalling. This study therefore examined the potential role of NO in nitration of tyrosine during the ripening of California sweet pepper.

METHODS

The NO content of green and red pepper fruit was determined spectrofluorometrically. Fruits at the breaking point between green and red coloration were incubated in the presence of NO for 1 h and then left to ripen for 3 d. Profiles of nitrated proteins were determined using an antibody against nitro-tyrosine (NO2-Tyr), and profiles of nitrosothiols were determined by confocal laser scanning microscopy. Nitrated proteins were identified by 2-D electrophoresis and MALDI-TOF/TOF analysis.

KEY RESULTS

Treatment with NO delayed the ripening of fruit. An enhancement of nitrosothiols and nitroproteins was observed in fruit during ripening, and this was reversed by the addition of exogenous NO gas. Six nitrated proteins were identified and were characterized as being involved in redox, protein, carbohydrate and oxidative metabolism, and in glutamate biosynthesis. Catalase was the most abundant nitrated protein found in both green and red fruit.

CONCLUSIONS

The RNS profile reported here indicates that ripening of pepper fruit is characterized by an enhancement of S-nitrosothiols and protein tyrosine nitration. The nitrated proteins identified have important functions in photosynthesis, generation of NADPH, proteolysis, amino acid biosynthesis and oxidative metabolism. The decrease of catalase in red fruit implies a lower capacity to scavenge H2O2, which would promote lipid peroxidation, as has already been reported in ripe pepper fruit.

Authors+Show Affiliations

Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, CSIC, Apartado 419, 18008 Granada, Spain and.Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, CSIC, Apartado 419, 18008 Granada, Spain and.Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, CSIC, Apartado 419, 18008 Granada, Spain and.Group of Biochemistry and Cell Signaling in Nitric Oxide. Department of Biochemistry and Molecular Biology, University of Jaén, 23071 Jaén, Spain.Group of Biochemistry and Cell Signaling in Nitric Oxide. Department of Biochemistry and Molecular Biology, University of Jaén, 23071 Jaén, Spain.Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, CSIC, Apartado 419, 18008 Granada, Spain and.Group of Antioxidants, Free Radicals and Nitric Oxide in Biotechnology, Food and Agriculture, Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, CSIC, Apartado 419, 18008 Granada, Spain and josemanuel.palma@eez.csic.es.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25814060

Citation

Chaki, Mounira, et al. "Ripening of Pepper (Capsicum Annuum) Fruit Is Characterized By an Enhancement of Protein Tyrosine Nitration." Annals of Botany, vol. 116, no. 4, 2015, pp. 637-47.
Chaki M, Álvarez de Morales P, Ruiz C, et al. Ripening of pepper (Capsicum annuum) fruit is characterized by an enhancement of protein tyrosine nitration. Ann Bot. 2015;116(4):637-47.
Chaki, M., Álvarez de Morales, P., Ruiz, C., Begara-Morales, J. C., Barroso, J. B., Corpas, F. J., & Palma, J. M. (2015). Ripening of pepper (Capsicum annuum) fruit is characterized by an enhancement of protein tyrosine nitration. Annals of Botany, 116(4), 637-47. https://doi.org/10.1093/aob/mcv016
Chaki M, et al. Ripening of Pepper (Capsicum Annuum) Fruit Is Characterized By an Enhancement of Protein Tyrosine Nitration. Ann Bot. 2015;116(4):637-47. PubMed PMID: 25814060.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ripening of pepper (Capsicum annuum) fruit is characterized by an enhancement of protein tyrosine nitration. AU - Chaki,Mounira, AU - Álvarez de Morales,Paz, AU - Ruiz,Carmelo, AU - Begara-Morales,Juan C, AU - Barroso,Juan B, AU - Corpas,Francisco J, AU - Palma,José M, Y1 - 2015/03/26/ PY - 2014/11/06/received PY - 2015/01/05/accepted PY - 2015/3/28/entrez PY - 2015/3/31/pubmed PY - 2016/6/18/medline KW - Capsicum annuum KW - MALDI-TOF/TOF KW - NO gas KW - RNS KW - Solanaceae KW - confocal laser scanning microscopy KW - nitric oxide KW - nitro-tyrosine KW - nitrosothiols KW - pepper fruit KW - protein nitration KW - proteomics KW - reactive nitrogen species KW - ripening SP - 637 EP - 47 JF - Annals of botany JO - Ann Bot VL - 116 IS - 4 N2 - BACKGROUND AND AIMS: Pepper (Capsicum annuum, Solanaceae) fruits are consumed worldwide and are of great economic importance. In most species ripening is characterized by important visual and metabolic changes, the latter including emission of volatile organic compounds associated with respiration, destruction of chlorophylls, synthesis of new pigments (red/yellow carotenoids plus xanthophylls and anthocyanins), formation of pectins and protein synthesis. The involvement of nitric oxide (NO) in fruit ripening has been established, but more work is needed to detail the metabolic networks involving NO and other reactive nitrogen species (RNS) in the process. It has been reported that RNS can mediate post-translational modifications of proteins, which can modulate physiological processes through mechanisms of cellular signalling. This study therefore examined the potential role of NO in nitration of tyrosine during the ripening of California sweet pepper. METHODS: The NO content of green and red pepper fruit was determined spectrofluorometrically. Fruits at the breaking point between green and red coloration were incubated in the presence of NO for 1 h and then left to ripen for 3 d. Profiles of nitrated proteins were determined using an antibody against nitro-tyrosine (NO2-Tyr), and profiles of nitrosothiols were determined by confocal laser scanning microscopy. Nitrated proteins were identified by 2-D electrophoresis and MALDI-TOF/TOF analysis. KEY RESULTS: Treatment with NO delayed the ripening of fruit. An enhancement of nitrosothiols and nitroproteins was observed in fruit during ripening, and this was reversed by the addition of exogenous NO gas. Six nitrated proteins were identified and were characterized as being involved in redox, protein, carbohydrate and oxidative metabolism, and in glutamate biosynthesis. Catalase was the most abundant nitrated protein found in both green and red fruit. CONCLUSIONS: The RNS profile reported here indicates that ripening of pepper fruit is characterized by an enhancement of S-nitrosothiols and protein tyrosine nitration. The nitrated proteins identified have important functions in photosynthesis, generation of NADPH, proteolysis, amino acid biosynthesis and oxidative metabolism. The decrease of catalase in red fruit implies a lower capacity to scavenge H2O2, which would promote lipid peroxidation, as has already been reported in ripe pepper fruit. SN - 1095-8290 UR - https://www.unboundmedicine.com/medline/citation/25814060/Ripening_of_pepper__Capsicum_annuum__fruit_is_characterized_by_an_enhancement_of_protein_tyrosine_nitration_ L2 - https://academic.oup.com/aob/article-lookup/doi/10.1093/aob/mcv016 DB - PRIME DP - Unbound Medicine ER -