Cell Fractionation.

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.

DOI: 10.1007/978-1-4939-7033-9_3

Cite this paper

@article{Petiti2017CellF, title={Cell Fractionation.}, author={Melissa Petiti and Laetitia Houot and Denis Duch{\'e}}, journal={Methods in molecular biology}, year={2017}, volume={1615}, pages={59-64} }