The grab-and-drop protocol: a novel strategy for membrane protein isolation and reconstitution from single cells.
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.
Department of Nanobiotechnology, University of Natural Resources and Life Sciences, Muthgasse 11, Vienna, 1190 Austria. o.ces@imperial.ac.uk.
Angelika
A
Schrems
John
J
Phillips
Duncan
D
Casey
Douglas
D
Wylie
Mira
M
Novakova
Uwe B
UB
Sleytr
David R
DR
Klug
David
D
Klug
Mark A A
MA
Neil
Bernhard
B
Schuster
Oscar
O
Ces
D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry*; D003110:Colonic Neoplasms / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000302:isolation & purification; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification*; D059010:Single-Cell Analysis / Q000295:instrumentation*; D018631:ras Proteins / Q000302:isolation & purification
D008565:Membrane Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D018631:ras Proteins