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Originally published In Press as doi:10.1074/mcp.M700002-MCP200 on June 14, 2007.
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Molecular & Cellular Proteomics 6:1459-1472, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.


Research

Salt-induced Changes in the Plasma Membrane Proteome of the Halotolerant Alga Dunaliella salina as Revealed by Blue Native Gel Electrophoresis and Nano-LC-MS/MS Analysis*,S

Adriana Katz{ddagger},§, Patrice Waridel§,||, Andrej Shevchenko|| and Uri Pick{ddagger}

From the {ddagger} Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel and || Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany

The halotolerant alga Dunaliella salina is a recognized model photosynthetic organism for studying plant adaptation to high salinity. The adaptation mechanisms involve major changes in the proteome composition associated with energy metabolism and carbon and iron acquisition. To clarify the molecular basis for the remarkable resistance to high salt, we performed a comprehensive proteomics analysis of the plasma membrane. Plasma membrane proteins were recognized by tagging intact cells with a membrane-impermeable biotin derivative. Proteins were resolved by two-dimensional blue native/SDS-PAGE and identified by nano-LC-MS/MS. Of 55 identified proteins, about 60% were integral membrane or membrane-associated proteins. We identified novel surface coat proteins, lipid-metabolizing enzymes, a new family of membrane proteins of unknown function, ion transporters, small GTP-binding proteins, and heat shock proteins. The abundance of 20 protein spots increased and that of two protein spots decreased under high salt. The major salt-regulated proteins were implicated in protein and membrane structure stabilization and within signal transduction pathways. The migration profiles of native protein complexes on blue native gels revealed oligomerization or co-migration of major surface-exposed proteins, which may indicate mechanisms of stabilization at high salinity.


To whom correspondence should be addressed. Tel.: 972-8-9342731; Fax: 972-8-9344118; E-mail: adriana.katz{at}weizmann.ac.il


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Copyright © 2007 by the American Society for Biochemistry and Molecular Biology.