title: Isoform-specific early trafficking of AMPA receptor flip and flop variants.
abstract: Flip and flop splice variants of AMPA receptor subunits are expressed in distinct but partly overlapping patterns and impart different desensitization kinetics to cognate receptor channels. In the absence of specific antibodies, isoform-specific differences in trafficking or localization of native flip and flop subunits remain uncharacterized. We report that in several transfected cell lines, transport of homomeric glutamate receptor (GluR)-D(flop) receptors is largely blocked at the endoplasmic reticulum (ER) exit, whereas GluR-D(flip) undergoes complex glycosylation and reaches the plasma membrane at >10x higher levels than GluR-D(flop), as determined by immunofluorescence, patch-clamp recordings and biochemical assays. The transport difference between flip and flop is independent of activity, is primarily determined by amino acid residue 780 (Leu in flop, Val in flip), and is manifested even in the secretion of the soluble ligand-binding domain, suggesting it is independent of oligomerization. Coexpression with stargazin or with the flip isoform rescues the surface expression of GluR-D(flop) near to the level exhibited by GluR-D(flip). Our results demonstrate that the extracellular flip/flop region, via interactions with ER luminal splice form-specific protein(s), plays a hitherto unappreciated and important role in AMPA-receptor trafficking.
authors.affiliation: Department of Biological and Environmental Sciences, Division of Biochemistry, Viikki Biocenter, University of Helsinki, FIN-00014 Helsinki, Finland.
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Coleman
authors.forename: Tommi
authors.initials: T
authors.lastname: Möykkynen
authors.forename: Chunlin
authors.initials: C
authors.lastname: Cai
authors.forename: Lotta
authors.initials: L
authors.lastname: von Ossowski
authors.forename: Esa
authors.initials: E
authors.lastname: Kuismanen
authors.forename: Esa R
authors.initials: ER
authors.lastname: Korpi
authors.forename: Kari
authors.initials: K
authors.lastname: Keinänen
mesh_terms: D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004274:DNA, Recombinant / Q000145:classification / Q000235:genetics / Q000378:metabolism; D014644:Genetic Variation / Q000502:physiology*; D006801:Humans; D051379:Mice; D020033:Protein Isoforms / Q000145:classification / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D018091:Receptors, AMPA / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D017470:Receptors, Glutamate / Q000145:classification / Q000235:genetics / Q000378:metabolism
chemical_list: D004274:DNA, Recombinant; D020033:Protein Isoforms; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; C109433:glutamate receptor type D