Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis.

Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis. Research Abstract Details 

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Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis. Abstract Text:

Blue native gel electrophoresis (BNGE) is a powerful tool for analyzing native protein complexes from biological membranes as well as water-soluble proteins. It can be used for determining relative molecular masses of protein complexes and their subunit composition and for the detection of subcomplexes. We describe the analysis by BNGE of in vitro import reactions composed of radiolabeled precursor proteins and isolated mitochondria. Such an analysis is a powerful tool to follow import intermediates and to study assembly of protein complexes. Analysis of import reactions by BNGE provides information on the molecular mass of the complex with which the imported precursor is associated. In addition, components of such a complex can be identified by incubating the mitochondrial lysate with either soluble antibodies or antibodies coupled to protein A matrix. The binding of soluble antibodies to specific complexes results in an observed shift in their apparent molecular mass (antibody shift). Alternatively, addition of matrix-bound antibodies followed by removal of the matrix from the mixture will result in depletion of the specific complex from the mitochondrial lysate (antibody depletion). The experimental details of these techniques are described.

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Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Methods in molecular biology (Clifton, N.J.)

VOLUME: 372

Page Numbers: 287-95

Journal Abbreviation: Methods Mol. Biol.

ISSN: 1064-3745

DAY: 4

MONTH: 03

YEAR: 2007

Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis. Information

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LANGUAGE: eng

NlmUniqueID: 9214969

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Grant and Affiliation Information for Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis.

AFFILIATION: Institut für Physiologische Chemie der Ludwig-Maximilians-Universität München, Germany.

Country: United States

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MEDLINETA: Methods Mol Biol

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