First published online December 23, 2004; 10.1104/pp.104.053041
Plant Physiology 137:104-116 (2005)
© 2005 American Society of Plant Biologists
CELL BIOLOGY AND SIGNAL TRANSDUCTION
Analysis of Detergent-Resistant Membranes in Arabidopsis. Evidence for Plasma Membrane Lipid Rafts1
Georg H.H. Borner2,
D. Janine Sherrier3,
Thilo Weimar,
Louise V. Michaelson,
Nathan D. Hawkins,
Andrew MacAskill,
Johnathan A. Napier,
Michael H. Beale,
Kathryn S. Lilley and
Paul Dupree*
Department of Biochemistry (G.H.H.B., D.J.S., T.W., A.M., P.D.) and Cambridge Centre for Proteomics (P.D., K.S.L.), University of Cambridge, Cambridge CB2 1QW, United Kingdom; and Crop Performance and Improvement, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom (L.V.M., N.D.H., J.A.N., M.H.B.)
The trafficking and function of cell surface proteins in eukaryotic cells may require association with detergent-resistant sphingolipid- and sterol-rich membrane domains. The aim of this work was to obtain evidence for lipid domain phenomena in plant membranes. A protocol to prepare Triton X-100 detergent-resistant membranes (DRMs) was developed using Arabidopsis (Arabidopsis thaliana) callus membranes. A comparative proteomics approach using two-dimensional difference gel electrophoresis and liquid chromatography-tandem mass spectrometry revealed that the DRMs were highly enriched in specific proteins. They included eight glycosylphosphatidylinositol-anchored proteins, several plasma membrane (PM) ATPases, multidrug resistance proteins, and proteins of the stomatin/prohibitin/hypersensitive response family, suggesting that the DRMs originated from PM domains. We also identified a plant homolog of flotillin, a major mammalian DRM protein, suggesting a conserved role for this protein in lipid domain phenomena in eukaryotic cells. Lipid analysis by gas chromatography-mass spectrometry showed that the DRMs had a 4-fold higher sterol-to-protein content than the average for Arabidopsis membranes. The DRMs were also 5-fold increased in sphingolipid-to-protein ratio. Our results indicate that the preparation of DRMs can yield a very specific set of membrane proteins and suggest that the PM contains phytosterol and sphingolipid-rich lipid domains with a specialized protein composition. Our results also suggest a conserved role of lipid modification in targeting proteins to both the intracellular and extracellular leaflet of these domains. The proteins associated with these domains provide important new experimental avenues into understanding plant cell polarity and cell surface processes.
1 This work was supported by the Biotechnology and Biological Sciences Research Council, by the Biotechnology and Biological Sciences Research Council Investigating Gene Function Initiative GARNet, by a European Community's Framework V Research Training Network Contract (HPRNCT200200262) Biointeractions (to T.W.), by the Nuffield Foundation, and by the Studienstiftung des Deutschen Volkes (scholarship to G.H.H.B.).
2 Present address: Cambridge Institute for Medical Research, Wellcome Trust/MRC Building, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2XY, UK.
3 Present address: Department of Plant and Soil Sciences and Delaware Biotechnology Institute, University of Delaware, Newark, DE 19711.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.053041.
* Corresponding author; e-mail p.dupree{at}bioc.cam.ac.uk; fax 441223333345.
Received September 6, 2004;
returned for revision October 17, 2004;
accepted October 23, 2004.
Related articles in Plant Physiol.:
- On the Inside
- Peter V. Minorsky
Plant Physiol. 2005 137: 1-2.
[Full Text]
This article has been cited by other articles:

|
 |

|
 |
 
E. Onelli, C. Prescianotto-Baschong, M. Caccianiga, and A. Moscatelli
Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold
J. Exp. Bot.,
August 1, 2008;
59(11):
3051 - 3068.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. Yalovsky, D. Bloch, N. Sorek, and B. Kost
Regulation of Membrane Trafficking, Cytoskeleton Dynamics, and Cell Polarity by ROP/RAC GTPases
Plant Physiology,
August 1, 2008;
147(4):
1527 - 1543.
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Chen, J. E. Markham, C. R. Dietrich, J. G. Jaworski, and E. B. Cahoon
Sphingolipid Long-Chain Base Hydroxylation Is Important for Growth and Regulation of Sphingolipid Content and Composition in Arabidopsis
PLANT CELL,
July 1, 2008;
20(7):
1862 - 1878.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. Mashiguchi, E. Urakami, M. Hasegawa, K. Sanmiya, I. Matsumoto, I. Yamaguchi, T. Asami, and Y. Suzuki
Defense-Related Signaling by Interaction of Arabinogalactan Proteins and {beta}-Glucosyl Yariv Reagent Inhibits Gibberellin Signaling in Barley Aleurone Cells
Plant Cell Physiol.,
February 1, 2008;
49(2):
178 - 190.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. Marmagne, M. Ferro, T. Meinnel, C. Bruley, L. Kuhn, J. Garin, H. Barbier-Brygoo, and G. Ephritikhine
A High Content in Lipid-modified Peripheral Proteins and Integral Receptor Kinases Features in the Arabidopsis Plasma Membrane Proteome
Mol. Cell. Proteomics,
November 1, 2007;
6(11):
1980 - 1996.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. Raffaele, S. Mongrand, P. Gamas, A. Niebel, and T. Ott
Genome-Wide Annotation of Remorins, a Plant-Specific Protein Family: Evolutionary and Functional Perspectives
Plant Physiology,
November 1, 2007;
145(3):
593 - 600.
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
N. Takata, J. Kasuga, D. Takezawa, K. Arakawa, and S. Fujikawa
Gene expression associated with increased supercooling capability in xylem parenchyma cells of larch (Larix kaempferi)
J. Exp. Bot.,
October 1, 2007;
58(13):
3731 - 3742.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
P. R. Ryan, Q. Liu, P. Sperling, B. Dong, S. Franke, and E. Delhaize
A Higher Plant {Delta}8 Sphingolipid Desaturase with a Preference for (Z)-Isomer Formation Confers Aluminum Tolerance to Yeast and Plants
Plant Physiology,
August 1, 2007;
144(4):
1968 - 1977.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
B. Lefebvre, F. Furt, M.-A. Hartmann, L. V. Michaelson, J.-P. Carde, F. Sargueil-Boiron, M. Rossignol, J. A. Napier, J. Cullimore, J.-J. Bessoule, et al.
Characterization of Lipid Rafts from Medicago truncatula Root Plasma Membranes: A Proteomic Study Reveals the Presence of a Raft-Associated Redox System
Plant Physiology,
May 1, 2007;
144(1):
402 - 418.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
N. Sorek, L. Poraty, H. Sternberg, E. Bar, E. Lewinsohn, and S. Yalovsky
Activation Status-Coupled Transient S Acylation Determines Membrane Partitioning of a Plant Rho-Related GTPase
Mol. Cell. Biol.,
March 15, 2007;
27(6):
2144 - 2154.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
A. K. Grennan
Lipid Rafts in Plants
Plant Physiology,
March 1, 2007;
143(3):
1083 - 1085.
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
P. Moreau, F. Brandizzi, S. Hanton, L. Chatre, S. Melser, C. Hawes, and B. Satiat-Jeunemaitre
The plant ER-Golgi interface: a highly structured and dynamic membrane complex
J. Exp. Bot.,
January 1, 2007;
58(1):
49 - 64.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Laloi, A.-M. Perret, L. Chatre, S. Melser, C. Cantrel, M.-N. Vaultier, A. Zachowski, K. Bathany, J.-M. Schmitter, M. Vallet, et al.
Insights into the Role of Specific Lipids in the Formation and Delivery of Lipid Microdomains to the Plasma Membrane of Plant Cells
Plant Physiology,
January 1, 2007;
143(1):
461 - 472.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Chen, G. Han, C. R. Dietrich, T. M. Dunn, and E. B. Cahoon
The Essential Nature of Sphingolipids in Plants as Revealed by the Functional Identification and Characterization of the Arabidopsis LCB1 Subunit of Serine Palmitoyltransferase
PLANT CELL,
December 1, 2006;
18(12):
3576 - 3593.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
H. S. Sardar, J. Yang, and A. M. Showalter
Molecular Interactions of Arabinogalactan Proteins with Cortical Microtubules and F-Actin in Bright Yellow-2 Tobacco Cultured Cells
Plant Physiology,
December 1, 2006;
142(4):
1469 - 1479.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. E. Markham, J. Li, E. B. Cahoon, and J. G. Jaworski
Separation and Identification of Major Plant Sphingolipid Classes from Leaves
J. Biol. Chem.,
August 11, 2006;
281(32):
22684 - 22694.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. J. Nelson, A. D. Hegeman, A. C. Harms, and M. R. Sussman
A Quantitative Analysis of Arabidopsis Plasma Membrane Using Trypsin-catalyzed 18O Labeling
Mol. Cell. Proteomics,
August 1, 2006;
5(8):
1382 - 1395.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J. Morel, S. Claverol, S. Mongrand, F. Furt, J. Fromentin, J.-J. Bessoule, J.-P. Blein, and F. Simon-Plas
Proteomics of Plant Detergent-resistant Membranes
Mol. Cell. Proteomics,
August 1, 2006;
5(8):
1396 - 1411.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. Grossmann, M. Opekarova, L. Novakova, J. Stolz, and W. Tanner
Lipid Raft-Based Membrane Compartmentation of a Plant Transport Protein Expressed in Saccharomyces cerevisiae
Eukaryot. Cell,
June 1, 2006;
5(6):
945 - 953.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. S Lilley and P. Dupree
Methods of quantitative proteomics and their application to plant organelle characterization
J. Exp. Bot.,
April 1, 2006;
57(7):
1493 - 1499.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. Baginsky and W. Gruissem
Arabidopsis thaliana proteomics: from proteome to genome
J. Exp. Bot.,
April 1, 2006;
57(7):
1485 - 1491.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
J.-U. Sutter, P. Campanoni, M. Tyrrell, and M. R. Blatt
Selective Mobility and Sensitivity to SNAREs Is Exhibited by the Arabidopsis KAT1 K+ Channel at the Plasma Membrane
PLANT CELL,
April 1, 2006;
18(4):
935 - 954.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
K. Terasaka, J. J. Blakeslee, B. Titapiwatanakun, W. A. Peer, A. Bandyopadhyay, S. N. Makam, O. R. Lee, E. L. Richards, A. S. Murphy, F. Sato, et al.
PGP4, an ATP Binding Cassette P-Glycoprotein, Catalyzes Auxin Transport in Arabidopsis thaliana Roots
PLANT CELL,
November 1, 2005;
17(11):
2922 - 2939.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
F. Roudier, A. G. Fernandez, M. Fujita, R. Himmelspach, G. H.H. Borner, G. Schindelman, S. Song, T. I. Baskin, P. Dupree, G. O. Wasteneys, et al.
COBRA, an Arabidopsis Extracellular Glycosyl-Phosphatidyl Inositol-Anchored Protein, Specifically Controls Highly Anisotropic Expansion through Its Involvement in Cellulose Microfibril Orientation
PLANT CELL,
June 1, 2005;
17(6):
1749 - 1763.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
H. Zheng, O. Rowland, and L. Kunst
Disruptions of the Arabidopsis Enoyl-CoA Reductase Gene Reveal an Essential Role for Very-Long-Chain Fatty Acid Synthesis in Cell Expansion during Plant Morphogenesis
PLANT CELL,
May 1, 2005;
17(5):
1467 - 1481.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. S. Gillmor, W. Lukowitz, G. Brininstool, J. C. Sedbrook, T. Hamann, P. Poindexter, and C. Somerville
Glycosylphosphatidylinositol-Anchored Proteins Are Required for Cell Wall Synthesis and Morphogenesis in Arabidopsis
PLANT CELL,
April 1, 2005;
17(4):
1128 - 1140.
[Abstract]
[Full Text]
[PDF]
|
 |
|
|
|