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Published 9 May 2005. doi:10.1083/jcb.200501121
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 169, Number 3, 399-404
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 Molecular constituents of neuronal AMPA receptors

Yuko Fukata1, Anastassios V. Tzingounis2, Jonathan C. Trinidad3, Masaki Fukata1, Alma L. Burlingame3, Roger A. Nicoll2, and David S. Bredt1

1 Department of Physiology, University of California, San Francisco, San Francisco, CA 94143
2 Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94143
3 Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143

Correspondence to David S. Bredt: bredt{at}itsa.ucsf.edu


Abstract
Dynamic regulation of {alpha}-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) underlies aspects of synaptic plasticity. Although numerous AMPAR-interacting proteins have been identified, their quantitative and relative contributions to native AMPAR complexes remain unclear. Here, we quantitated protein interactions with neuronal AMPARs by immunoprecipitation from brain extracts. We found that stargazin-like transmembrane AMPAR regulatory proteins (TARPs) copurified with neuronal AMPARs, but we found negligible binding to GRIP, PICK1, NSF, or SAP-97. To facilitate purification of neuronal AMPAR complexes, we generated a transgenic mouse expressing an epitope-tagged GluR2 subunit of AMPARs. Taking advantage of this powerful new tool, we isolated two populations of GluR2 containing AMPARs: an immature complex with the endoplasmic reticulum chaperone immunoglobulin-binding protein and a mature complex containing GluR1, TARPs, and PSD-95. These studies establish TARPs as the auxiliary components of neuronal AMPARs.

Y. Fukata and M. Fukata's present address is Laboratory of Genomics and Proteomics, National Institute for Longevity Sciences, Obu, Aichi 474-8522, Japan.

Abbreviations used in this paper: AMPA, {alpha}-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid; AMPAR, AMPA receptor; CBP, calmodulin-binding peptide; IAP, immunoaffinity purification; LTD, long-term depression; NMDA, N-methyl-D-aspartate; TARP, transmembrane AMPAR regulatory protein.


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