Atomic force microscopy of cloned nicotinic acetylcholine receptor expressed in Xenopus oocytes.
The nicotinic acetylcholine receptor (AChR) was expressed in Xenopus oocytes from in vitro transcribed mRNA and was imaged by atomic force microscopy. A characteristic pentameric structure of AChR was readily observed on the extracellular face of the cell membrane, with a central pore surrounded by protruding AChR subunits. These structures were seen only in mRNA-injected oocytes that also gave acetylcholine-induced membrane currents. The size of individual AChR channels, the angles between subunits, and the interchannel spacing were all compatible with the current model of AChR. In addition, localized patches of microscopic AChR clustering were observed, with packing density approaching that at the neuromuscular junction. These findings show the potential of studying cloned membrane proteins in oocytes for both their surface topography and their structure-function relationship in native membrane without the need for crystallization.
Lal
R
R
Department of Medicine, University of Chicago, IL 60637.
Yu
L
L
D000818:Animals; D005260:Female; D066298:In Vitro Techniques; D007473:Ion Channels / Q000648:ultrastructure*; D051379:Mice; D008853:Microscopy / Q000379:methods*; D009865:Oocytes / Q000648:ultrastructure; D011978:Receptors, Nicotinic / Q000648:ultrastructure*; D011994:Recombinant Proteins; D014982:Xenopus laevis
D007473:Ion Channels; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins