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Defining Membrane Protein Localization by Isopycnic Density Gradients

  • Rhys A. Dunstan
  • Iain D. Hay
  • Trevor Lithgow
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1615)

Abstract

In many bacteria, membrane proteins account for around one-third of the proteome and can represent much more than half of the mass of a membrane. Classic techniques in cell biology can be applied to characterise bacterial membranes and their membrane protein constituents. Here we describe a protocol for the purification of outer and inner membranes from Escherichia coli. The procedure can be applied with minor modifications to other bacterial species, including those carrying capsular polysaccharide attached to the outer membrane.

Key words

Sucrose density gradient Membrane biogenesis Beta-barrel proteins Cytoplasmic membrane Lipoproteins 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Rhys A. Dunstan
    • 1
  • Iain D. Hay
    • 1
  • Trevor Lithgow
    • 1
  1. 1.Department of Microbiology and Infection and Immunity Program, Biomedicine Discovery InstituteMonash UniversityMelbourneAustralia

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