Skip to main content
SpringerLink
Log in

Evaluation of Lipids for the Study of Photosynthetic Membranes

  • Protocol
  • First Online:
Photosynthesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1770))

Abstract

The biological role of lipids goes far beyond the formation of a structural membrane bilayer platform for membrane proteins and controlling fluxes across the membranes. For example, in photosynthetic thylakoid membranes, lipids occupy well-defined binding niches within protein complexes and determine the structural organization of membrane proteins and their function by controlling generic physicochemical membrane properties. In this chapter, two-dimensional thin-layer chromatography (2D TLC) and gas chromatography (GC) techniques are presented for quantitative analysis of lipid classes and fatty acids in thylakoid membranes. In addition, lipid extraction methods from isolated thylakoid membranes and leaves are described together with a procedure for derivatization of fatty acids to fatty acid methyl esters (FAME) that is required for GC analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Boudière L, Michaud M, Petroutsos D, Rébeillé F, Falconet D, Bastien O, Roy S, Finazzi G, Rolland N, Jouhet J, Block MA, Maréchal E (2014) Glycerolipids in photosynthesis: composition, synthesis and trafficking. Biochim Biophys Acta 1837:470–480

    Article  CAS  PubMed  Google Scholar 

  2. Block MA, Dorne AJ, Joyard J, Douce R (1983) Preparation and characterization of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts. 2. Biochemical characterization. J Biol Chem 258:3281–3286

    Google Scholar 

  3. Murata N, Siegenthalter PA (1998) Chapter 1: Lipids in photosynthesis: an overview. In: Murata N, Siegenthaler PA (eds) Lipids in photosynthesis: structure, function and genetics. Kluwer Academic Publishers, Dordrecht, pp 1–20

    Google Scholar 

  4. Garab G, Ughy B, Goss R (2016) Role of MGDG and non-bilayer lipid phases in the structure and dynamics of chloroplast thylakoid membranes. Subcell Biochem 86:127–157

    Article  CAS  PubMed  Google Scholar 

  5. Christie WW, Dobson G (1999) Thin-layer chromatography re-visited. Lipid Technol 11:64–66

    CAS  Google Scholar 

  6. Wever V, Doermann P, Hoelz G (2013) Chapter 8: Analysis and quantification of plant membrane lipids by thin-layer chromatography and gas chromatography. In: Munnik T, Heilmann I (eds) Plant lipid signaling protocols. Springer Science and Business Media, p 69–78

    Google Scholar 

  7. Wang Z, Benning C (2011) Arabidopsis thaliana polar glycerolipid profiling by thin layer chromatography (TLC) coupled with gas-liquid chromatography (GLC). J Vis Exp 49:2518

    Google Scholar 

  8. Wu Z, Zhang Q, Li N, Pu Y, Wang B, Zhang T (2017) Comparison of critical methods developed for fatty acid analysis: a review. J Sep Sci 40:288–298

    Article  CAS  PubMed  Google Scholar 

  9. Porra RJ, Thompson WA, Kriedermann PE (1989) Determination of accurate extinction coefficient and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim Biophys Acta 975:384–394

    Article  CAS  Google Scholar 

  10. Christie WW (1989) Chapter 5: Gas chromatographic analysis of fatty acid derivatives. In: Christi WW (ed) Gas chromatography and lipids. Oily Press Ltd, Dundee, pp 73–75

    Google Scholar 

Download references

Acknowledgments

The authors acknowledge support from the US National Science Foundation (MCB-1616982), the US Department of Energy (DE-SC0017160), the US Department of Agriculture (ARC grant WNP00775), and the US National Institute of Food and Agriculture (NIFA, 2011-68005-30416) and USDA National Institute of Food and Agriculture, Hatch project 0119.

Author information

Authors and Affiliations

  1. Institute of Biological Chemistry, Washington State University, Pullman, WA, USA

    Helmut Kirchhoff & Robert Yarbrough

Authors
  1. Helmut Kirchhoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert Yarbrough
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Helmut Kirchhoff .

Editor information

Editors and Affiliations

  1. Niceville, Florida, USA

    Sarah Covshoff

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kirchhoff, H., Yarbrough, R. (2018). Evaluation of Lipids for the Study of Photosynthetic Membranes. In: Covshoff, S. (eds) Photosynthesis. Methods in Molecular Biology, vol 1770. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7786-4_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7786-4_18

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7785-7

  • Online ISBN: 978-1-4939-7786-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation