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JXB Advance Access originally published online on June 11, 2008
Journal of Experimental Botany 2008 59(11):2979-2990; doi:10.1093/jxb/ern156
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© The Author [2008]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Grape berry plasma membrane proteome analysis and its differential expression during ripening

Jiangwei Zhang1 *, Huiqin Ma2 *, Jidong Feng3, Lei Zeng1, Zeng Wang2 and Shangwu Chen1,{dagger}

1College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China
2College of Agriculture and Biotechnology, China Agricultural University, Beijing, China
3College of Biological Sciences, China Agricultural University, Beijing, China

{dagger} To whom correspondence should be addressed. E-mail: swchen{at}cau.edu.cn

High purity berry plasma membranes (PMs) of Vitis vinifera L. cv. Cabernet Sauvignon were isolated by two-phase partitioning of microsome fractions at different stages of berry ripening. PM proteins resolvable by the detergent cocktail of CHAPS and ASB-14 were separated by two-dimensional electrophoresis. A total of 119 protein spots from pre-véraison berry PMs on 2-D gels detected with silver staining were subjected to MALDI-TOF mass spectrometry analysis. Sixty-two spots were identified as putative PM proteins, with 1–6 predicted transmembrane helices, including true PM proteins such as ATP synthase, ABC transporters, and GTP-binding proteins reported in plants. They were then grouped into eight functional categories, mainly involved in transport, metabolism, signal transduction, and protein synthesis. Another 11 spots were identified as proteins of unknown function. The véraison and post-véraison samples stained 98 and 86 spots on the gels, respectively. During the berry ripening process, total PM protein content gradually decreased. Among all identified proteins, 12 showed significant differences in terms of their relative abundance. Increasing ubiquitin proteolysis and cytoskeleton proteins were observed from pre-véraison to post-véraison. Zeatin O-glucosyltransferase peaked at véraison, while ubiquitin-conjugating enzyme E2-21 was down-regulated at this stage. This proteome research provides the first information on PM protein characterization during the grape berry ripening process.

Key words: Grape berry, MALDI-TOF-MS, plasma membrane, proteomics, two-dimensional electrophoresis

* These authors contributed equally to this work.

Received 14 March 2008; Revised 4 May 2008 Accepted 6 May 2008


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