Chapter title |
Expression and Purification of the Individual Bam Components BamB–E
|
---|---|
Chapter number | 14 |
Book title |
The BAM Complex
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2871-2_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2870-5, 978-1-4939-2871-2
|
Authors |
Suraaj Aulakh, Kelly H. Kim, Mark Paetzel, Aulakh, Suraaj, Kim, Kelly H., Paetzel, Mark |
Abstract |
BamB, BamC, BamD, and BamE are lipoproteins that, along with the integral membrane protein BamA, form the β-barrel assembly machinery (BAM) complex in the outer-membrane of Gram-negative bacteria. Elucidating the roles that these lipoproteins play in the β-barrel assembly process requires both structural and functional studies that rely on milligram quantities of pure protein. Here, we describe a simple protocol for expressing individual BamB-BamE proteins in Escherichia coli and purifying them by nickel affinity and size-exclusion chromatography. This protocol yields pure proteins in amounts that are sufficient for crystallization trials, in vitro protein-protein interaction studies, NMR, and other biochemical experiments. |
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Geographical breakdown
Country | Count | As % |
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Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Unspecified | 1 | 20% |
Librarian | 1 | 20% |
Student > Ph. D. Student | 1 | 20% |
Unknown | 2 | 40% |
Readers by discipline | Count | As % |
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Unspecified | 1 | 20% |
Biochemistry, Genetics and Molecular Biology | 1 | 20% |
Computer Science | 1 | 20% |
Unknown | 2 | 40% |