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Plant and Cell Physiology Advance Access originally published online on December 23, 2008
Plant and Cell Physiology 2009 50(2):341-359; doi:10.1093/pcp/pcn202
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Alterations in Detergent-Resistant Plasma Membrane Microdomains in Arabidopsis thaliana During Cold Acclimation

Anzu Minami1, Masayuki Fujiwara2, Akari Furuto3, Yoichiro Fukao2, Tetsuro Yamashita4, Masaharu Kamo5, Yukio Kawamura1 and Matsuo Uemura1,3,*

1The 21st Century Center of Excellence Program, Iwate University, Morioka, 020-8550 Japan
2Laboratory of Plant Molecular and Plant Protein Analysis, Plant Science Education Unit, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630-0101 Japan
3Cryobiofrontier Research Center, Faculty of Agriculture, Iwate University, Morioka, 020-8550 Japan
4Department of Biological Chemistry and Food Sciences, Faculty of Agriculture, Iwate University, Morioka, 020-8550 Japan
5Department of Biochemistry, Iwate Medical University School of Dentistry, Morioka, 020-8505 Japan

*Corresponding author: E-mail, uemura{at}iwate-u.ac.jp; Fax, +81-19-621-6253.


  Abstract

Microdomains in the plasma membrane (PM) have been proposed to be involved in many important cellular events in plant cells. To understand the role of PM microdomains in plant cold acclimation, we isolated the microdomains as detergent-resistant plasma membrane fractions (DRMs) from Arabidopsis seedlings and compared lipid and protein compositions before and after cold acclimation. The DRM was enriched in sterols and glucocerebrosides, and the proportion of free sterols in the DRM increased after cold acclimation. The protein-to-lipid ratio in the DRM was greater than that in the total PM fraction. The protein amount recovered in DRMs decreased gradually during cold acclimation. Cold acclimation further resulted in quantitative changes in DRM protein profiles. Subsequent mass spectrometry and Western blot analyses revealed that P-type H+-ATPases, aquaporins and endocytosis-related proteins increased and, conversely, tubulins, actins and V-type H+-ATPase subunits decreased in DRMs during cold acclimation. Functional categorization of cold-responsive proteins in DRMs suggests that plant PM microdomains function as platforms of membrane transport, membrane trafficking and cytoskeleton interaction. These comprehensive changes in microdomains may be associated with cold acclimation of Arabidopsis.

Keywords: Arabidopsis - Cold acclimation - Detergent-resistant membrane - 2D-DIGE - Microdomains - Plasma membrane

Abbreviations: ASG, acylated sterylglucoside; BSA, bovine serum albumin; 2D-DIGE, two-dimensional difference gel electrophoresis; DRM, detergent-resistant plasma membrane fraction; DTT, dithiothreitol; GlcCer, glucocerebroside; FS, free sterol; LC-MS/MS, liquid chromatography-mass spectrometry/mass spectrometry; MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; PLD, phospholipase D; PL, phospholipid; PM, plasma membrane; P-ATPase, P-type H+-ATPase; SG, sterylglucoside; TLC, thin-layer chromatography; V-ATPase, V-type H+-ATPase

(Received November 5, 2008; Accepted December 16, 2008)
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