title: Protein composition of immunoprecipitated synaptic ribbons.
abstract: The synaptic ribbon is an electron-dense structure found in hair cells and photoreceptors. The ribbon is surrounded by neurotransmitter-filled vesicles and considered to play a role in vesicle release. We generated an objective, quantitative analysis of the protein composition of the ribbon complex using a mass spectrometry-based proteomics analysis. Our use of affinity-purified ribbons and control IgG immunoprecipitations ensure that the identified proteins are indeed associated with the ribbon complex. The use of mouse tissue, where the proteome is complete, generated a comprehensive analysis of the candidates. We identified 30 proteins (comprising 56 isoforms and subunits) associated with the ribbon complex. The ribbon complex primarily comprises proteins found in conventional synapses, which we categorized into 6 functional groups: vesicle handling (38.5%), scaffold (7.3%), cytoskeletal molecules (20.6%), phosphorylation enzymes (10.6%), molecular chaperones (8.2%), and transmembrane proteins from the presynaptic membrane firmly attached to the ribbon (11.3%). The 3 CtBP isoforms represent the major protein in the ribbon whether calculated by molar amount (30%) or by mass (20%). The relatively high quantity of phosphorylation enzymes suggests a very active and regulated structure. The ribbon appears to comprise a concentrated cluster of proteins dealing with vesicle creation, retention and distribution, and consequent exocytosis.
authors: Eaton-Peabody Laboratory, Department of Otology and Laryngology, Massachusetts Eye and Ear Infirmary and Harvard Medical School, 243 Charles Street, Boston, Massachusetts 02114, United States.
authors: A
authors: A
authors: Kantardzhieva
authors: M
authors: M
authors: Peppi
authors: W S
authors: WS
authors: Lane
authors: W F
authors: WF
authors: Sewell
mesh_terms: D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000737:chemistry; D005089:Exocytosis; D007074:Immunoglobulin G / Q000378:metabolism; D007150:Immunohistochemistry; D047468:Immunoprecipitation / Q000379:methods*; D013058:Mass Spectrometry; D051379:Mice; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000145:classification / Q000378:metabolism; D010786:Photoreceptor Cells / Q000737:chemistry*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D012160:Retina / Q000166:cytology; D013569:Synapses / Q000737:chemistry*
chemical_list: D007074:Immunoglobulin G; D009419:Nerve Tissue Proteins; D020543:Proteome