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Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
Published on: August 14, 2011
An assay to monitor the membrane integration of single-span proteins
Katrin Krumpe1, Doron Rapaport
1Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany.
Abstract:
In vitro import experiments with isolated organelles are a powerful tool for investigation of the biogenesis of proteins. A key issue in such experiments is an assay to distinguish between correctly and incorrectly imported proteins. Here we describe an assay to monitor in vitro the proper membrane integration of single-span proteins. In this assay non-imported proteins are distinguished from correctly imported protein species by labelling of unprotected cysteine residues and a resulting migration shift in SDS-PAGE.
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