The grab-and-drop protocol: A novel strategy for membrane protein isolation and reconstitution from single cells

Angelika Schrems, John Phillips, Duncan Casey, Douglas Wylie, Mira Novakova, Uwe B. Sleytr, David Klug, Mark A.A. Neil, Bernhard Schuster, Oscar Ces*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

7 Citations (Scopus)

Abstract

We present a rapid and robust technique for the sampling of membrane-associated proteins from the surface of a single, live cell and their subsequent deposition onto a solid-supported lipid bilayer. As a proof of principle, this method has been used to extract green fluorescent protein (EGFP) labelled K-ras proteins located at the inner leaflet of the plasma membrane of colon carcinoma cells and to transfer them to an S-layer supported lipid bilayer system. The technique is non-destructive, meaning that both the cell and proteins are intact after the sampling operation, offering the potential for repeated measurements of the same cell of interest. This system provides the ideal tool for the investigation of cellular heterogeneity, as well as a platform for the investigation of rare cell types such as circulating tumour cells.

Original languageEnglish
Pages (from-to)3296-3304
Number of pages9
JournalAnalyst
Volume139
Issue number13
Early online date3 Jun 2014
DOIs
Publication statusPublished - 7 Jul 2014

Keywords

  • Cell Line, Tumor
  • Cell Membrane
  • Colonic Neoplasms
  • Green Fluorescent Proteins
  • Humans
  • Membrane Proteins
  • Single-Cell Analysis
  • ras Proteins
  • Journal Article
  • Research Support, Non-U.S. Gov't

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