Chapter title |
Cell Fractionation
|
---|---|
Chapter number | 3 |
Book title |
Bacterial Protein Secretion Systems
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7033-9_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7031-5, 978-1-4939-7033-9
|
Authors |
Melissa Petiti, Laetitia Houot, Denis Duché |
Editors |
Laure Journet, Eric Cascales |
Abstract |
Protein function is generally dependent on its subcellular localisation. In Gram-negative bacteria such as Escherichia coli, a protein can be targeted to five different compartments: the cytoplasm, the inner membrane, the periplasm, the outer membrane and the extracellular medium. Different approaches can be used to determine the protein localisation within a cell such as in silico identification of protein signal sequences and motifs, electron microscopy and immunogold labelling, optical fluorescence microscopy, and biochemical technics. In this chapter, we describe a simple and efficient method to isolate the different compartments of Escherichia coli by a fractionation method and to determine the presence of the protein of interest. For inner membrane proteins we propose a method to discriminate between integral and peripheral membrane proteins. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 14 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 3 | 21% |
Student > Bachelor | 3 | 21% |
Researcher | 2 | 14% |
Student > Master | 2 | 14% |
Lecturer | 1 | 7% |
Other | 1 | 7% |
Unknown | 2 | 14% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 9 | 64% |
Agricultural and Biological Sciences | 1 | 7% |
Immunology and Microbiology | 1 | 7% |
Chemistry | 1 | 7% |
Unknown | 2 | 14% |