title: Structural basis of PI(4,5)P2-dependent regulation of GluA1 by phosphatidylinositol-5-phosphate 4-kinase, type II, alpha (PIP5K2A).
abstract: Ionotropic glutamate receptors are the most important excitatory receptors in the central nervous system, and their impairment can lead to multiple neuronal diseases. Here, we show that glutamate-induced currents in oocytes expressing GluA1 are increased by coexpression of the schizophrenia-associated phosphoinositide kinase PIP5K2A. This effect was due to enhanced membrane abundance and was blunted by a point mutation (N251S) in PIP5K2A. An increase in GluA1 currents was also observed upon acute injection of PI(4,5)P2, the main product of PIP5K2A. By expression of wild-type and mutant PIP5K2A in human embryonic kidney cells, we were able to provide evidence of impaired kinase activity of the mutant PIP5K2A. We defined the region K813-K823 of GluA1 as critical for the PI(4,5)P2 effect by performing an alanine scan that suggested PI(4,5)P2 binding to this area. A PIP strip assay revealed PI(4,5)P2 binding to the C-terminal GluA1 peptide. The present observations disclose a novel mechanism in the regulation of GluA1.
authors.affiliation: Institute for Genetics of Heart Diseases (IfGH), -Myocellular Electrophysiology, Department of Cardiovascular Medicine, University Hospital Muenster, 48149, Muenster, Germany.
authors.forename: Guiscard
authors.initials: G
authors.lastname: Seebohm
authors.forename: Eva
authors.initials: E
authors.lastname: Wrobel
authors.forename: Michael
authors.initials: M
authors.lastname: Pusch
authors.forename: Markus
authors.initials: M
authors.lastname: Dicks
authors.forename: Jan
authors.initials: J
authors.lastname: Terhag
authors.forename: Veronika
authors.initials: V
authors.lastname: Matschke
authors.forename: Ina
authors.initials: I
authors.lastname: Rothenberg
authors.forename: Oana N
authors.initials: ON
authors.lastname: Ursu
authors.forename: Fabian
authors.initials: F
authors.lastname: Hertel
authors.forename: Lutz
authors.initials: L
authors.lastname: Pott
authors.forename: Florian
authors.initials: F
authors.lastname: Lang
authors.forename: Eric
authors.initials: E
authors.lastname: Schulze-Bahr
authors.forename: Michael
authors.initials: M
authors.lastname: Hollmann
authors.forename: Raphael
authors.initials: R
authors.lastname: Stoll
authors.forename: Nathalie
authors.initials: N
authors.lastname: Strutz-Seebohm
mesh_terms: D000409:Alanine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D018091:Receptors, AMPA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014981:Xenopus
chemical_list: D000409:Alanine; D019269:Phosphatidylinositol 4,5-Diphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); D018091:Receptors, AMPA; C514871:PIP4K2A protein, human