Nature Methods
- 3, 1007 - 1012 (2006)
Published online: 22 October 2006; | doi:10.1038/nmeth965
Two-chamber AFM: probing membrane proteins separating two aqueous compartmentsRui Pedro Gonçalves1, Guillaume Agnus2, Pierre Sens3, Christine Houssin4, Bernard Bartenlian2 & Simon Scheuring11
Institut Curie, UMR168-CNRS, 26 Rue d'Ulm, 75248 Paris, France. 2
IEF, MMS, Université Paris-Sud, Bat. 220, 91405 Orsay, France. 3
ESPCI, CNRS-UMR 7083, 10 rue Vauquelin, 75231 Paris, France. 4
LBMII, IGM, Université Paris-Sud, Bat. 360, 91405 Orsay, France.
Correspondence should be addressed to Simon Scheuring simon.scheuring@curie.fr Biological membranes compartmentalize and define physical borders of cells. They are crowded with membrane proteins that fulfill diverse crucial functions. About one-third of all genes in organisms code for, and the majority of drugs target, membrane proteins. To combine structure and function analysis of membrane proteins, we designed a two-chamber atomic force microscopy (AFM) setup that allows investigation of membranes spanned over nanowells, therefore separating two aqueous chambers. We imaged nonsupported surface layers (S layers) of Corynebacterium glutamicum at sufficient resolution to delineate a 15 Å–wide protein pore. We probed the elastic and yield moduli of nonsupported membranes, giving access to the lateral interaction energy between proteins. We combined AFM and fluorescence microscopy to demonstrate the functionality of proteins in the setup by documenting proton pumping by Halobacterium salinarium purple membranes.
MORE ARTICLES LIKE THIS These links to content published by NPG are automatically generated.
|
