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Expression of mouse-Torpedo acetylcholine receptor subunit chimeras and hybrids in Xenopus oocytes

Mayne, Katharine Mixter and Yoshii, Kiyonori and Yu, Lei and Lester, Henry A. and Davidson, Norman (1987) Expression of mouse-Torpedo acetylcholine receptor subunit chimeras and hybrids in Xenopus oocytes. Molecular Brain Research, 2 (3). pp. 191-197. ISSN 0169-328X. https://resolver.caltech.edu/CaltechAUTHORS:20201013-140644515

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Abstract

In this study, in vitro synthesized mRNA encoding mouse and Torpedo nicotinic acetylcholine receptor subunits was injected into Xenopus oocytes, followed by assays for assembly onto the oocyte surface (using [¹²⁵I]α-bungarotoxin binding) and for acetylcholine-induced conductances (using voltage clamp). We constructed hybrid acetylcholine receptors in Xenopus oocytes by injecting all 8 possible combinations of 4 subunit-specific mRNAs in which a single subunit is derived from the other species. For each hybrid combination, there is detectable assembly and conductance. We also constructed cDNA clones that encode chimeric acetylcholine receptor subunits in which part of the γ subunit from Torpedo was replaced by the homologous region of the δ subunit from mouse. None of the chimeric subunits was able to replace the Torpedo γ, mouse δ, or Torpedo δ subunit with regard to assembly or function. We therefore conclude that widely spaced (and unknown) parts of the protein chain are required for the intersubunit interactions that eventually lead to functional assembly of the receptor.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/0169-328x(87)90026-xDOIArticle
ORCID:
AuthorORCID
Lester, Henry A.0000-0002-5470-5255
Additional Information:© 1987 Published by Elsevier. Accepted 5 May 1987. We thank Drs. T. Claudio, S. Heinemann, and D. Noonan for kindly providing Torpedo AChR cDNA clones, Drs. D. Mead and D. Melton for providing SP6 expression plasmids and information concerning the SP6 transcription system, Drs. J. Jackson and E. Barnard for the chick a cDNA clone, Dr. J.P. Merlie for the mouse a cDNA clone, and Drs. R. Stroud and J. Finer-Moore for helpful discussions. This work has been supported in part by research grants GM-10991 and NS-11756 from the National Institutes of Health, a research grant for the Muscular Dystrophy Association, support from the government of Japan for K.Y., Predoctoral Training Grant GM-07616 from the National Institutes of Health to K.M.M., and a California Foundation for Biochemical Research Fellowship to L.Y.
Funders:
Funding AgencyGrant Number
NIHGM-10991
NIHNS-11756
Muscular Dystrophy AssociationUNSPECIFIED
Ministry of Education, Culture, Sports, Science and Technology (MEXT)UNSPECIFIED
NIH Predoctoral FellowshipGM-07616
California Foundation for Biochemical ResearchUNSPECIFIED
Subject Keywords:In vitro transcription; Ion channel
Issue or Number:3
Record Number:CaltechAUTHORS:20201013-140644515
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201013-140644515
Official Citation:Katharine Mixter Mayne, Kiyonori Yoshii, Lei Yu, Henry A. Lester, Norman Davidson, Expression of mouse-Torpedo acetylcholine receptor subunit chimeras and hybrids in Xenopus oocytes, Molecular Brain Research, Volume 2, Issue 3, 1987, Pages 191-197, ISSN 0169-328X, https://doi.org/10.1016/0169-328X(87)90026-X.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:106038
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:13 Oct 2020 22:04
Last Modified:13 Oct 2020 22:04

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