title: RIBEYE(B)-domain binds to lipid components of synaptic vesicles in an NAD(H)-dependent, redox-sensitive manner.
abstract: Synaptic ribbons are needed for fast and continuous exocytosis in ribbon synapses. RIBEYE is a main protein component of synaptic ribbons and is necessary to build the synaptic ribbon. RIBEYE consists of a unique A-domain and a carboxyterminal B-domain, which binds NAD(H). Within the presynaptic terminal, the synaptic ribbons are in physical contact with large numbers of synaptic vesicle (SV)s. How this physical contact between ribbons and synaptic vesicles is established at a molecular level is not well understood. In the present study, we demonstrate that the RIBEYE(B)-domain can directly interact with lipid components of SVs using two different sedimentation assays with liposomes of defined chemical composition. Similar binding results were obtained with a SV-containing membrane fraction. The binding of liposomes to RIBEYE(B) depends upon the presence of a small amount of lysophospholipids present in the liposomes. Interestingly, binding of liposomes to RIBEYE(B) depends on NAD(H) in a redox-sensitive manner. The binding is enhanced by NADH, the reduced form, and is inhibited by NAD+, the oxidized form. Lipid-mediated attachment of vesicles is probably part of a multi-step process that also involves additional, protein-dependent processes.
authors.affiliation: Institute of Anatomy and Cell Biology, Department of Neuroanatomy, Saarland University, Medical School Homburg/Saar, Homburg/Saar 66421, Germany dr.karin.schwarz@uks.eu frank.schmitz@uks.eu.
authors.forename: Karin
authors.initials: K
authors.lastname: Schwarz
authors.affiliation: Institute of Anatomy and Cell Biology, Department of Neuroanatomy, Saarland University, Medical School Homburg/Saar, Homburg/Saar 66421, Germany dr.karin.schwarz@uks.eu frank.schmitz@uks.eu.
authors.forename: Frank
authors.initials: F
authors.lastname: Schmitz
mesh_terms: D000818:Animals; D002417:Cattle; D002725:Chloroform; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D000432:Methanol; D009243:NAD / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012160:Retina / Q000737:chemistry / Q000378:metabolism*; D012997:Solvents; D013569:Synapses / Q000737:chemistry / Q000378:metabolism*; D013572:Synaptic Vesicles / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005136:Eye Proteins; D008081:Liposomes; D010743:Phospholipids; D011993:Recombinant Fusion Proteins; D012997:Solvents; D009243:NAD; D002725:Chloroform; D002784:Cholesterol; D000432:Methanol
keywords: NAD(H); RIBEYE; liposome; redox-sensitivity; ribbon synapse; synaptic ribbon