title: Nicotinamide adenine dinucleotide-dependent binding of the neuronal Ca2+ sensor protein GCAP2 to photoreceptor synaptic ribbons.
abstract: Guanylate cyclase activating protein 2 (GCAP2) is a recoverin-like Ca2+-sensor protein known to modulate guanylate cyclase activity in photoreceptor outer segments. GCAP2 is also present in photoreceptor ribbon synapses where its function is unknown. Synaptic ribbons are active zone-associated presynaptic structures in the tonically active photoreceptor ribbon synapses and contain RIBEYE as a unique and major protein component. In the present study, we demonstrate by various independent approaches that GCAP2 specifically interacts with RIBEYE in photoreceptor synapses. We show that the flexible hinge 2 linker region of RIBEYE(B) domain that connects the nicotinamide adenine dinucleotide (NADH)-binding subdomain with the substrate-binding subdomain (SBD) binds to the C terminus of GCAP2. We demonstrate that the RIBEYE-GCAP2 interaction is induced by the binding of NADH to RIBEYE. RIBEYE-GCAP2 interaction is modulated by the SBD. GCAP2 is strongly expressed in synaptic terminals of light-adapted photoreceptors where GCAP2 is found close to synaptic ribbons as judged by confocal microscopy and proximity ligation assays. Virus-mediated overexpression of GCAP2 in photoreceptor synaptic terminals leads to a reduction in the number of synaptic ribbons. Therefore, GCAP2 is a prime candidate for mediating Ca2+-dependent dynamic changes of synaptic ribbons in photoreceptor synapses.
authors.affiliation: Department of Neuroanatomy, Institute for Anatomy and Cell Biology, Saarland University Medical School, 66421 Homburg/Saar, Germany.
authors.forename: Jagadeesh Kumar
authors.initials: JK
authors.lastname: Venkatesan
authors.forename: Sivaraman
authors.initials: S
authors.lastname: Natarajan
authors.forename: Karin
authors.initials: K
authors.lastname: Schwarz
authors.forename: Sabine I
authors.initials: SI
authors.lastname: Mayer
authors.forename: Kannan
authors.initials: K
authors.lastname: Alpadi
authors.forename: Venkat Giri
authors.initials: VG
authors.lastname: Magupalli
authors.forename: Ching-Hwa
authors.initials: CH
authors.lastname: Sung
authors.forename: Frank
authors.initials: F
authors.lastname: Schmitz
mesh_terms: D000429:Alcohol Oxidoreductases; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D056970:Co-Repressor Proteins; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051604:Guanylate Cyclase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D009243:NAD / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D012160:Retina / Q000378:metabolism; D013569:Synapses / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D056970:Co-Repressor Proteins; D004268:DNA-Binding Proteins; D051604:Guanylate Cyclase-Activating Proteins; C496740:Guca1b protein, mouse; D010750:Phosphoproteins; D009243:NAD; D000429:Alcohol Oxidoreductases; C111341:Ctbp2 protein, mouse
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