SpringerProtocols: Abstract: Membrane Protein Insertion and Secretion in Bacteria

Membrane Protein Insertion and Secretion in Bacteria

By: Jeanine de Keyzer^{2 3}, Martin van der Laan^{2 4}, Arnold J.M. Driessen²

Abstract

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Export of secretory proteins across and insertion of membrane proteins into the cytoplasmic membrane of Escherichia coli and other bacteria is mediated by the enzyme complex translocase. The last decade has seen a major advance in the understanding of the mechanism of these processes. A large part of this progress can be attributed to the development of general and powerful methods to study the translocase activity in vitro. Here we describe a transcription–translation method used to analyze the insertion of membrane proteins into E. coli inner membrane vesicles and a rapid and quantitative fluorescent method to analyze the translocation of secretory proteins.

Affiliation(s): (2) Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute and the Materials Science Center Plus, University of Groningen, The Netherlands
(3) Faculty of Life Sciences, Michael Smith Building, University of Manchester, Manchester, UK
(4) Institut für Biochemie und Molekularbiologie, Universität Freiburg, Hermann-Herder-Straβe 7, Freiburg, Germany

Book Title: Protein Targeting Protocols

Series: Methods in Molecular Biology | Volume: 390 | Pub. Date: May-21-2007 | Page Range: 17-32 | DOI: 10.1007/978-1-59745-466-7_2

Subject: Protein Science

Key Words: E. coli translocase - protein translocation - membrane protein insertion - transcription - translation - fluorescent imaging - SecA - SecYEG.

References

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