Title | Counting bungarotoxin binding sites of nicotinic acetylcholine receptors in mammalian cells with high signal/noise ratios. |
Publication Type | Journal Article |
Year of Publication | 2010 |
Authors | Simonson PD, Deberg HA, Ge P, Alexander JK, Jeyifous O, Green WN, Selvin PR |
Journal | Biophys J |
Volume | 99 |
Issue | 10 |
Pagination | L81-3 |
Date Published | 2010 Nov 17 |
ISSN | 1542-0086 |
Keywords | alpha7 Nicotinic Acetylcholine Receptor, Binding Sites, Bungarotoxins, Fluorescent Dyes, HEK293 Cells, Humans, Neuromuscular Junction, Photobleaching, Receptors, Nicotinic |
Abstract | Nicotinic acetylcholine receptors are some of the most studied synaptic proteins; however, many questions remain that can only be answered using single molecule approaches. Here we report our results from single α7 and neuromuscular junction type nicotinic acetylcholine receptors in mammalian cell membranes. By labeling the receptors with fluorophore-labeled bungarotoxin, we can image individual receptors and count the number of bungarotoxin-binding sites in receptors expressed in HEK 293 cells. Our results indicate that there are two bungarotoxin-binding sites in neuromuscular junction receptors, as expected, and five in α7 receptors, clarifying previous uncertainty. This demonstrates a valuable technique for counting subunits in membrane-bound proteins at the single molecule level, with nonspecialized optics and with higher signal/noise ratios than previous fluorescent protein-based techniques. |
DOI | 10.1016/j.bpj.2010.08.076 |
Alternate Journal | Biophys J |
PubMed ID | 21081055 |
Grant List | GM086214 / GM / NIGMS NIH HHS / United States DA019695 / DA / NIDA NIH HHS / United States NS043782 / NS / NINDS NIH HHS / United States |
Related Faculty:
Paul Simonson, M.D., Ph.D.