Chapter title |
An In Vitro Assay for Outer Membrane Protein Assembly by the BAM Complex
|
---|---|
Chapter number | 16 |
Book title |
The BAM Complex
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2871-2_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2870-5, 978-1-4939-2871-2
|
Authors |
Giselle Roman-Hernandez, Harris D. Bernstein, Roman-Hernandez, Giselle, Bernstein, Harris D. |
Abstract |
To elucidate the mechanism of a biochemical process it is often essential to reconstitute the reaction in vitro using the minimal set of factors required to drive the reaction to completion. Here, we describe a method to reconstitute the folding and membrane integration of bacterial outer membrane (OM) proteins that have a characteristic β-barrel structure. In this method the BAM complex, a heteroligomer that catalyzes the membrane integration of β-barrel proteins, is first purified and inserted into small lipid vesicles. Denatured OM proteins are then assembled and integrated into the vesicles in the presence of a molecular chaperone called SurA. |
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Unknown | 3 | 17% |