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Quantitative Assessment of the Chloroplast Lipidome

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Plastids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1829))

Abstract

In plants and algae, photosynthetic membranes have a unique lipid composition. They differ from all other cellular membranes by their very low amount of phospholipids, besides some phosphatidylglycerol (PG), and high proportion of glycolipids. These glycolipids are the uncharged galactolipids, i.e., monogalactosyldiacylglycerol and digalactosyldiacylglycerol (MGDG and DGDG), and an anionic sulfolipid, i.e., sulfoquinovosyldiacylglycerol (SQDG). In all photosynthetic membranes analyzed to date, from cyanobacteria to algae, protists, and plants, the lipid quartet constituted by MGDG, DGDG, SQDG, and PG has been highly conserved but the composition in fatty acids of these lipids can vary a lot from an organism to another. To better understand chloroplast biogenesis, it is therefore essential to know their lipid content. Establishing chloroplast lipidome requires first to purify chloroplast from plant or algae tissue. Here we describe the methods to extract lipids, quantify the lipids of the chloroplast, and qualify and quantify the different lipid classes that might be present in these fractions.

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Reference

  1. Boudière L, Michaud M, Petroutsos D et al (2014) Glycerolipids in photosynthesis: composition, synthesis and trafficking. Biochim Biophys Acta Bioenerg 1837(4):470–480

    Article  CAS  Google Scholar 

  2. Domingues P, Amado FML, Santana-Marquesa MGO et al (1998) Constant neutral loss scanning for the characterization of glycerol phosphatidylcholine phospholipids. J Am Soc Mass Spectrom 9(11):1189–1195

    Article  CAS  Google Scholar 

  3. Brügger B, Erben G, Sandhoff R et al (1997) Quantitative analysis of biological membrane lipids at the low picomole level by nano-electrospray ionization tandem mass spectrometry. Proc Natl Acad Sci U S A 94(6):2339–2344

    Article  PubMed  PubMed Central  Google Scholar 

  4. Taguchi R, Houjou T, Nakanishi H et al (2005) Focused lipidomics by tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 823(1):26–36

    Article  CAS  PubMed  Google Scholar 

  5. Hsu FF, Turk J (2000) Characterization of phosphatidylinositol, phosphatidylinositol-4-phosphate, and phosphatidylinositol-4,5-bisphosphate by electrospray ionization tandem mass spectrometry: a mechanistic study. J Am Soc Mass Spectrom 11(11):986–999

    Article  CAS  PubMed  Google Scholar 

  6. Li-Beisson Y, Shorrosh B, Beisson F et al (2010) Acyl-lipid metabolism. Arabidopsis Book 8:e0133

    Article  PubMed  PubMed Central  Google Scholar 

  7. Welti R, Wang X, Williams TD (2003) Electrospray ionization tandem mass spectrometry scan modes for plant chloroplast lipids. Anal Biochem 314(1):149–152

    Article  CAS  PubMed  Google Scholar 

  8. Naumann I, Klein BC, Bartel SJ et al (2011) Identification of sulfoquinovosyldiacyglycerides from Phaeodactylum tricornutum by matrix-assisted laser desorption/ionization QTrap time-of-flight hybrid mass spectrometry. Rapid Commun Mass Spectrom 25(17):2517–2523

    Article  CAS  PubMed  Google Scholar 

  9. Moreau RA, Doehlert DC, Welti R et al (2008) The identification of mono-, di-, tri-, and tetragalactosyl-diacylglycerols and their natural estolides in oat kernels. Lipids 43(6):533–548

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Benning C, Huang ZH, Gage DA (1995) Accumulation of a novel glycolipid and a betaine lipid in cells of Rhodobacter sphaeroides grown under phosphate limitation. Arch Biochem Biophys 317(1):103–111

    Article  CAS  PubMed  Google Scholar 

  11. Hsu FF, Turk J (2003) Electrospray ionization/tandem quadrupole mass spectrometric studies on phosphatidylcholines: the fragmentation processes. J Am Soc Mass Spectrom 14(4):352–363

    Article  CAS  PubMed  Google Scholar 

  12. Hsu FF, Turk J (2000) Characterization of phosphatidylethanolamine as a lithiated adduct by triple quadrupole tandem mass spectrometry with electrospray ionization. J Mass Spectrom 35(5):595–606

    Article  CAS  PubMed  Google Scholar 

  13. Hsu FF, Turk J (2001) Studies on phosphatidylglycerol with triple quadrupole tandem mass spectrometry with electrospray ionization: fragmentation processes and structural characterization. J Am Soc Mass Spectrom 12(9):1036–1043

    Article  CAS  Google Scholar 

  14. Zianni R, Bianco G, Lelario F et al (2013) Fatty acid neutral losses observed in tandem mass spectrometry with collision-induced dissociation allows regiochemical assignment of sulfoquinovosyl-diacylglycerols. J Mass Spectrom 48(2):205–215

    Article  CAS  PubMed  Google Scholar 

  15. Guella G, Frassanito R, Mancini I (2003) A new solution for an old problem: the regiochemical distribution of the acyl chains in galactolipids can be established by electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom 17(17):1982–1994

    Article  CAS  PubMed  Google Scholar 

  16. Abida H, Dolch LJ, Meï C et al (2015) Membrane glycerolipid remodeling triggered by nitrogen and phosphorus starvation in Phaeodactylum tricornutum. Plant Physiol 167(1):118–136

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by the French National Research Agency (ANR-13-ADAP-0008 Reglisse and ANR-10-LABEX-04 GRAL Labex, Grenoble Alliance for Integrated Structural Cell Biology).

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Correspondence to Juliette Jouhet .

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Gros, V., Jouhet, J. (2018). Quantitative Assessment of the Chloroplast Lipidome. In: Maréchal, E. (eds) Plastids. Methods in Molecular Biology, vol 1829. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8654-5_16

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  • DOI: https://doi.org/10.1007/978-1-4939-8654-5_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8653-8

  • Online ISBN: 978-1-4939-8654-5

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