title: Presynaptic control of glycine transporter 2 (GlyT2) by physical and functional association with plasma membrane Ca2+-ATPase (PMCA) and Na+-Ca2+ exchanger (NCX).
abstract: Fast inhibitory glycinergic transmission occurs in spinal cord, brainstem, and retina to modulate the processing of motor and sensory information. After synaptic vesicle fusion, glycine is recovered back to the presynaptic terminal by the neuronal glycine transporter 2 (GlyT2) to maintain quantal glycine content in synaptic vesicles. The loss of presynaptic GlyT2 drastically impairs the refilling of glycinergic synaptic vesicles and severely disrupts neurotransmission. Indeed, mutations in the gene encoding GlyT2 are the main presynaptic cause of hyperekplexia in humans. Here, we show a novel endogenous regulatory mechanism that can modulate GlyT2 activity based on a compartmentalized interaction between GlyT2, neuronal plasma membrane Ca(2+)-ATPase (PMCA) isoforms 2 and 3, and Na(+)/Ca(2+)-exchanger 1 (NCX1). This GlyT2·PMCA2,3·NCX1 complex is found in lipid raft subdomains where GlyT2 has been previously found to be fully active. We show that endogenous PMCA and NCX activities are necessary for GlyT2 activity and that this modulation depends on lipid raft integrity. Besides, we propose a model in which GlyT2·PMCA2-3·NCX complex would help Na(+)/K(+)-ATPase in controlling local Na(+) increases derived from GlyT2 activity after neurotransmitter release.
authors.affiliation: From the Department of Biochemistry, Weill Cornell Medical College, New York, New York 10065.
authors.forename: Jaime
authors.initials: J
authors.lastname: de Juan-Sanz
authors.affiliation: the Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049-Madrid, Spain, the Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, 46009-Valencia, Spain, the IdiPAZ-Hospital, Universitario La Paz, 28046-Madrid, Spain, and.
authors.forename: Enrique
authors.initials: E
authors.lastname: Núñez
authors.affiliation: the Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049-Madrid, Spain, the Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, 46009-Valencia, Spain, the IdiPAZ-Hospital, Universitario La Paz, 28046-Madrid, Spain, and.
authors.forename: Francisco
authors.initials: F
authors.lastname: Zafra
authors.affiliation: the Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006-Badajoz, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Berrocal
authors.affiliation: the Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006-Badajoz, Spain.
authors.forename: Isaac
authors.initials: I
authors.lastname: Corbacho
authors.affiliation: the Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049-Madrid, Spain, the Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, 46009-Valencia, Spain, the IdiPAZ-Hospital, Universitario La Paz, 28046-Madrid, Spain, and.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Ibáñez
authors.affiliation: the Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049-Madrid, Spain, the Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, 46009-Valencia, Spain, the IdiPAZ-Hospital, Universitario La Paz, 28046-Madrid, Spain, and.
authors.forename: Esther
authors.initials: E
authors.lastname: Arribas-González
authors.affiliation: the Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006-Badajoz, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Marcos
authors.affiliation: the Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049-Madrid, Spain, the Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, 46009-Valencia, Spain, the IdiPAZ-Hospital, Universitario La Paz, 28046-Madrid, Spain, and.
authors.forename: Beatriz
authors.initials: B
authors.lastname: López-Corcuera
authors.affiliation: the Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06006-Badajoz, Spain.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Mata
authors.affiliation: the Centro de Biología Molecular "Severo Ochoa," Universidad Autónoma de Madrid, Consejo Superior de Investigaciones Científicas, 28049-Madrid, Spain, the Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, 46009-Valencia, Spain, the IdiPAZ-Hospital, Universitario La Paz, 28046-Madrid, Spain, and caragon@cbm.csic.es.
authors.forename: Carmen
authors.initials: C
authors.lastname: Aragón
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D050487:Glycine Plasma Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D021962:Membrane Microdomains / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000187:drug effects; D061251:Primary Cell Culture; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D011984:Sensory Receptor Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009435:Synaptic Transmission; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: C101670:2-(2-(4-(4-nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate; D050487:Glycine Plasma Membrane Transport Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C493269:Slc6a5 protein, rat; D019831:Sodium-Calcium Exchanger; D047392:beta-Cyclodextrins; C423122:caloxin 2A1; C108732:methyl-beta-cyclodextrin; C110374:sodium-calcium exchanger 1; D053499:Plasma Membrane Calcium-Transporting ATPases; D013890:Thiourea
keywords: Calcium ATPase; GlyT2; Lipid Raft; Neurotransmitter Transport; Proteomics; Sodium-Calcium Exchange