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(Received for publication, January 26, 1996, and in revised form, April 30, 1996)
From the Nicotinic acetylcholine receptors formed from
combinations of
Volume 271, Number 30,
Issue of July 26, 1996
pp. 17656-17665
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
5 Subunits with
3, 
2, and 4 Subunits
,
,
,
,
Department of Neuroscience and
¶ Department of Pharmacology, University of Pennsylvania Medical
School, Philadelphia, Pennsylvania 19104-6074 and the '' Vision Science
Research Center, University of Alabama,
Birmingham, Alabama 35294-4390
3, 
2, 4, and 5 subunits are found in
chicken ciliary ganglion neurons and some human neuroblastoma cell
lines. We studied the co-expression of various combinations of cloned
human 3, 2, 4, and 5 subunits in Xenopus
oocytes. Expression on the surface membrane was found only for
combinations of 32, 34, 325, and 345
subunits but not for other combinations of one, two, or three of these
subunits. 5 subunits assembled inside the oocyte with 2 but not
with 3 subunits or other 5 subunits. 5 subunits coassembled
very efficiently with 32 or 34 combinations. The presence
of 5 subunits had very little effect on the binding affinities for
epibatidine of receptors containing also 3 and 2 or 3 and 4
subunits. The presence of 5 subunits increased the rate of
desensitization of both receptors containing also 3 and 2 or 3
and 4 subunits. In the case of receptors containing 3 and 4
subunits, the addition of 5 subunits had little effect on the
responses to acetylcholine or nicotine. However, in the case of
receptors containing 3 and 2 subunits, the addition of 5
subunits reduced the EC50 for acetylcholine from 28 to 0.5 µM and the EC50 for nicotine from 6.8 to 1.9 µM, while increasing the efficacy of nicotine from 50%
on 32 receptors to 100% on 325 receptors. Both 32
and 325 receptors expressed in oocytes sedimented at the same
11 S value as native 3-containing receptors from the human
neuroblastoma cell line SH-SY5Y. In the receptors from the
neuroblastoma 3, 2, and 5 subunits were co-assembled, and 56%
of the receptor subtypes containing 3 subunits also contained 2
subunits. The 2 subunit-containing receptors from SH-SY5Y cells
exhibited the high affinity for epibatidine characteristic of receptors
formed from 3 and 2 or 3, 2, and 5 subunits rather than
the low affinity exhibited by receptors formed from 3 and 4 or
3, 4, and 5 subunits. Nicotine, like the structurally similar
toxin epibatidine, also distinguishes by binding affinity two subtypes
of receptors containing 3 subunits in SH-SY5Y cells. The affinities
of 32 receptors expressed in oocytes were similar to the
affinities of native 3 containing receptors from SH-SY5Y cells
for acetylcholine, cytisine, and 1,1-dimethyl-4-phenylpiperazinium.
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C R. Yu and L. W Role Functional contribution of the {alpha}5 subunit to neuronal nicotinic channels expressed by chick sympathetic ganglion neurones J. Physiol., June 15, 1998; 509(3): 667 - 681. [Abstract] [Full Text] [PDF]
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W. G. Conroy and D. K. Berg Nicotinic Receptor Subtypes in the Developing Chick Brain: Appearance of a Species Containing the alpha 4, beta 2, and alpha 5 Gene Products Mol. Pharmacol., March 1, 1998; 53(3): 392 - 401. [Abstract] [Full Text]
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J. M. Kulak, T. A. Nguyen, B. M. Olivera, and J. M. McIntosh alpha -Conotoxin MII Blocks Nicotine-Stimulated Dopamine Release in Rat Striatal Synaptosomes J. Neurosci., July 15, 1997; 17(14): 5263 - 5270. [Abstract] [Full Text] [PDF]
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X. Peng, V. Gerzanich, R. Anand, F. Wang, and J. Lindstrom Chronic Nicotine Treatment Up-Regulates alpha 3 and alpha 7 Acetylcholine Receptor Subtypes Expressed by the Human Neuroblastoma Cell Line SH-SY5Y Mol. Pharmacol., May 1, 1997; 51(5): 776 - 784. [Abstract] [Full Text]
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M. Rogers, L. M. Colquhoun, J. W. Patrick, and J. A. Dani Calcium Flux Through Predominantly Independent Purinergic ATP and Nicotinic Acetylcholine Receptors J Neurophysiol, March 1, 1997; 77(3): 1407 - 1417. [Abstract] [Full Text] [PDF]
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 J.-P. Changeux, A. Bessis, J.-P. Bourgeois, P.-J. Corringer, A. Devillers-Thiery, J.-L. Eisele, M. Kerszberg, C. Lena, N. Le Novere, M. Picciotto, et al. Nicotinic Receptors and Brain Plasticity Cold Spring Harb Symp Quant Biol, January 1, 1996; 61(0): 343 - 362. [Abstract] [PDF]
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G. B. Wells, L. Lin, E. M. Jeanclos, and R. Anand Assembly and Ligand Binding Properties of the Water-soluble Extracellular Domains of the Glutamate Receptor 1 Subunit J. Biol. Chem., January 26, 2001; 276(5): 3031 - 3036. [Abstract] [Full Text] [PDF]
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