HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplemental data All Versions of this Article: T600018-MCP200v1 5/11/2185    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zischka, H. Articles by Ueffing, M. Search for Related Content PubMed PubMed Citation Articles by Zischka, H. Articles by Ueffing, M. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 5:2185-2200, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Technology

Differential Analysis of Saccharomyces cerevisiae Mitochondria by Free Flow Electrophoresis^*,S

Hans Zischka^,,¶, Ralf J. Braun, Enrico P. Marantidis, Dietmute Büringer^||, Carsten Bornhövd^**, Stefanie M. Hauck, Oliver Demmer, Christian J. Gloeckner, Andreas S. Reichert^**, Frank Madeo and Marius Ueffing^,

From the Institutes of Human Genetics and Toxicology, GSF-National Research Center for Environment and Health, 85764 Munich-Neuherberg, Germany, ^|| Department of Systems and Computational Neurobiology, Max Planck Institute for Neurobiology, 82152 Martinsried, Germany, ^** Adolf-Butenandt-Institute for Physiological Chemistry, Ludwig-Maximilians University Munich, 81377 Munich, Germany, Institute of Molecular Biosciences, University of Graz, Graz A-8010, Austria, and Institute for Human Genetics, Technical University Munich, 81675 Munich, Germany

One major problem concerning the electrophoresis of mitochondria is the heterogeneity of mitochondrial appearance especially under pathological conditions. We show here the use of zone electrophoresis in a free flow electrophoresis device (ZE-FFE) as an analytical sensor to discriminate between different yeast mitochondrial populations. Impairment of the structural properties of the organelles by hyperosmotic stress resulted in broad separation profiles. Conversely untreated mitochondria gave rise to homogeneous populations reflected by sharp separation profiles. Yeast mitochondria with altered respiratory activity accompanied by a different outer membrane proteome composition could be discriminated based on electrophoretic deflection. Proteolysis of the mitochondrial surface proteome and the deletion of a single major protein species of the mitochondrial outer membrane altered the ZE-FFE deflection of these organelles. To demonstrate the usefulness of ZE-FFE for the analysis of mitochondria associated with pathological processes, we analyzed mitochondrial fractions from an apoptotic yeast strain. The cdc48^S565G strain carries a mutation in the CDC48 gene that is an essential participant in the endoplasmic reticulum-associated protein degradation pathway. Mutant cells accumulate polyubiquitinated proteins in microsomal and mitochondrial extracts. Subsequent ZE-FFE characterization could distinguish a mitochondrial subfraction specifically enriched with polyubiquitinated proteins from the majority of non-affected mitochondria. This result demonstrates that ZE-FFE may give important information on the specific properties of subpopulations of a mitochondrial preparation allowing a further detailed functional analysis.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. Jamai, P. A. Salome, S. H. Schilling, A. P.M. Weber, and C. R. McClung Arabidopsis Photorespiratory Serine Hydroxymethyltransferase Activity Requires the Mitochondrial Accumulation of Ferredoxin-Dependent Glutamate Synthase PLANT CELL, February 1, 2009; 21(2): 595 - 606. [Abstract] [Full Text] [PDF]

				O. Drews, R. Wildgruber, C. Zong, U. Sukop, M. Nissum, G. Weber, A. V. Gomes, and P. Ping Mammalian Proteasome Subpopulations with Distinct Molecular Compositions and Proteolytic Activities Mol. Cell. Proteomics, November 1, 2007; 6(11): 2021 - 2031. [Abstract] [Full Text] [PDF]