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5. Purification and protein composition of oil bodies from Brassica napus seeds

Author

Jolivet Pascale, Tailliart Kathleen, Boulard Céline, Nesi Nathalie, and Chardot Thierry

Subjects
Brassica napus, Arabidopsis thaliana, oil bodies, oleosins, protein composition, Oils, fats, and waxes, TP670-699
Abstract

Seed oil bodies are intracellular particles to store lipids as food reserves in oleaginous plants. Description of oil body-associated proteins of Arabidopsis thaliana has been recently reported whereas only few data are available in the case of rapeseed. Oil bodies have been prepared from two double-low varieties of Brassica napus seeds, a standard variety (Explus) and an oleic variety (Cabriolet). Oil bodies have been purified using floatation technique in the successive presence of high salt concentration, detergent or urea in order to remove non-specifically trapped proteins. The integrity of the oil bodies has been verified and their size estimated. Their protein and fatty acid contents have been determined. The proteins composing these organelles were extracted, separated by denaturing gel electrophoresis, digested by trypsin and their peptides were subsequently analyzed by liquid chromatography-tandem mass spectrometry. Protein identification was performed using Arabidopsis thaliana protein sequence database and a collection of Expressed Sequence Tag (EST) of Brassica napus generated from the framework of the French plant genomics programme “Genoplante”. This led to the identification of a limited number of proteins: eight oleosins showing a high similarity each other and representing up to 75% of oil body proteins, a 11 β hydroxysteroid dehydrogenase-like protein highly homologous to the same protein from A. thaliana, and only few contaminating proteins associated with myrosinase activity.

Published
2006
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8. Design and Characterisation of a Randomized Food Intervention That Mimics Exposure to a Typical UK Diet to Provide Urine Samples for Identification and Validation of Metabolite Biomarkers of Food Intake

Author

Willis, Naomi D., primary, Lloyd, Amanda J., additional, Xie, Long, additional, Stiegler, Martina, additional, Tailliart, Kathleen, additional, Garcia-Perez, Isabel, additional, Chambers, Edward S., additional, Beckmann, Manfred, additional, Draper, John, additional, and Mathers, John C., additional

Published
2020
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9. A Standardized Strategy for Simultaneous Quantification of Urine Metabolites to Validate Development of a Biomarker Panel Allowing Comprehensive Assessment of Dietary Exposure

Author

Beckmann, Manfred, primary, Wilson, Thomas, additional, Zubair, Hassan, additional, Lloyd, Amanda J., additional, Lyons, Laura, additional, Phillips, Helen, additional, Tailliart, Kathleen, additional, Gregory, Nicholas, additional, Thatcher, Rhys, additional, Garcia‐Perez, Isabel, additional, Frost, Gary, additional, Mathers, John M., additional, and Draper, John, additional

Published
2020
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10. Developing community-based urine sampling methods to deploy biomarker technology for the assessment of dietary exposure

Author

Lloyd, Amanda J, primary, Wilson, Thomas, additional, Willis, Naomi D, additional, Lyons, Laura, additional, Phillips, Helen, additional, Janssen, Hayley G, additional, Stiegler, Martina, additional, Xie, Long, additional, Tailliart, Kathleen, additional, Beckmann, Manfred, additional, Stevenson, Leo, additional, Mathers, John C, additional, and Draper, John, additional

Published
2020
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11. Developing a Food Exposure and Urine Sampling Strategy for Dietary Exposure Biomarker Validation in Free‐Living Individuals

Author

Lloyd, Amanda J., primary, Willis, Naomi D., additional, Wilson, Thomas, additional, Zubair, Hassan, additional, Xie, Long, additional, Chambers, Edward, additional, Garcia‐Perez, Isabel, additional, Tailliart, Kathleen, additional, Beckmann, Manfred, additional, Mathers, John C., additional, and Draper, John, additional

Published
2019
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13. Purification and protein composition of oil bodies from Brassica napus seeds

Author

Jolivet, Pascale, Tailliart, Kathleen, Boulard, Céline, Nesi, Nathalie, Chardot, Thierry, Jolivet, Pascale, Tailliart, Kathleen, Boulard, Céline, Nesi, Nathalie, and Chardot, Thierry

Abstract

Seed oil bodies are intracellular particles to store lipids as food reserves in oleaginous plants. Description of oil body-associated proteins of Arabidopsis thaliana has been recently reported whereas only few data are available in the case of rapeseed. Oil bodies have been prepared from two double-low varieties of Brassica napus seeds, a standard variety (Explus) and an oleic variety (Cabriolet). Oil bodies have been purified using floatation technique in the successive presence of high salt concentration, detergent or urea in order to remove non-specifically trapped proteins. The integrity of the oil bodies has been verified and their size estimated. Their protein and fatty acid contents have been determined. The proteins composing these organelles were extracted, separated by denaturing gel electrophoresis, digested by trypsin and their peptides were subsequently analyzed by liquid chromatography-tandem mass spectrometry. Protein identification was performed using Arabidopsis thaliana protein sequence database and a collection of Expressed Sequence Tag (EST) of Brassica napus generated from the framework of the French plant genomics programme “Genoplante”. This led to the identification of a limited number of proteins: eight oleosins showing a high similarity each other and representing up to 75% of oil body proteins, a 11 β hydroxysteroid dehydrogenase-like protein highly homologous to the same protein from A. thaliana, and only few contaminating proteins associated with myrosinase activity.

Published
2006
Full Text
View/download PDF

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