title: Structural characterization of heparin-induced glyceraldehyde-3-phosphate dehydrogenase protofibrils preventing α-synuclein oligomeric species toxicity.
abstract: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional enzyme that has been associated with neurodegenerative diseases. GAPDH colocalizes with α-synuclein in amyloid aggregates in post-mortem tissue of patients with sporadic Parkinson disease and promotes the formation of Lewy body-like inclusions in cell culture. In a previous work, we showed that glycosaminoglycan-induced GAPDH prefibrillar species accelerate the conversion of α-synuclein to fibrils. However, it remains to be determined whether the interplay among glycosaminoglycans, GAPDH, and α-synuclein has a role in pathological states. Here, we demonstrate that the toxic effect exerted by α-synuclein oligomers in dopaminergic cell culture is abolished in the presence of GAPDH prefibrillar species. Structural analysis of prefibrillar GAPDH performed by small angle x-ray scattering showed a particle compatible with a protofibril. This protofibril is shaped as a cylinder 22 nm long and a cross-section diameter of 12 nm. Using biocomputational techniques, we obtained the first all-atom model of the GAPDH protofibril, which was validated by cross-linking coupled to mass spectrometry experiments. Because GAPDH can be secreted outside the cell where glycosaminoglycans are present, it seems plausible that GAPDH protofibrils could be assembled in the extracellular space kidnapping α-synuclein toxic oligomers. Thus, the role of GAPDH protofibrils in neuronal proteostasis must be considered. The data reported here could open alternative ways in the development of therapeutic strategies against synucleinopathies like Parkinson disease.
authors.affiliation: From the Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, and Instituto de Química Biológica "Dr. Bernabé Bloj," FBQF-UNT, Chacabuco 461, T4000ILI Tucumán, Argentina.
authors.forename: César L
authors.initials: CL
authors.lastname: Ávila
authors.affiliation: From the Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, and Instituto de Química Biológica "Dr. Bernabé Bloj," FBQF-UNT, Chacabuco 461, T4000ILI Tucumán, Argentina.
authors.forename: Clarisa M
authors.initials: CM
authors.lastname: Torres-Bugeau
authors.affiliation: the Instituto de Física da Universidade de São Paulo, Rua do Matão, Travessa R, 187, São Paulo, Brazil.
authors.forename: Leandro R S
authors.initials: LR
authors.lastname: Barbosa
authors.affiliation: the Instituto de Física da Universidade de São Paulo, Rua do Matão, Travessa R, 187, São Paulo, Brazil.
authors.forename: Elisa Morandé
authors.initials: EM
authors.lastname: Sales
authors.affiliation: INSERM U1127, CNRS UMR 7225, Institut de Cerveau et de la Moelle Epinière, Paris, France, the Laboratoire Croissance, Réparation et Régénération Tissulaires, CNRS EAC 7149, Université Paris Est Créteil, Université Paris Est, F-94000, Créteil, France, and.
authors.forename: Mohand O
authors.initials: MO
authors.lastname: Ouidja
authors.affiliation: INSERM U1127, CNRS UMR 7225, Institut de Cerveau et de la Moelle Epinière, Paris, France.
authors.forename: Sergio B
authors.initials: SB
authors.lastname: Socías
authors.affiliation: the Departamento de Química Biológica, Centro de Investigaciones en Química Biológica de Córdoba, CONICET, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Haya de la Torre y Medina Allende, Ciudad Universitaria, X5000HUA Córdoba, Argentina.
authors.forename: M Soledad
authors.initials: MS
authors.lastname: Celej
authors.affiliation: INSERM U1127, CNRS UMR 7225, Institut de Cerveau et de la Moelle Epinière, Paris, France.
authors.forename: Rita
authors.initials: R
authors.lastname: Raisman-Vozari
authors.affiliation: the Laboratoire Croissance, Réparation et Régénération Tissulaires, CNRS EAC 7149, Université Paris Est Créteil, Université Paris Est, F-94000, Créteil, France, and.
authors.forename: Dulce
authors.initials: D
authors.lastname: Papy-Garcia
authors.affiliation: the Instituto de Física da Universidade de São Paulo, Rua do Matão, Travessa R, 187, São Paulo, Brazil.
authors.forename: Rosangela
authors.initials: R
authors.lastname: Itri
authors.affiliation: From the Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, and Instituto de Química Biológica "Dr. Bernabé Bloj," FBQF-UNT, Chacabuco 461, T4000ILI Tucumán, Argentina, rosana@fbqf.unt.edu.ar.
authors.forename: Rosana N
authors.initials: RN
authors.lastname: Chehín
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000494:pharmacology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D018696:Neuroprotective Agents / Q000737:chemistry / Q000494:pharmacology; D055503:Protein Multimerization / Q000187:drug effects*; D017433:Protein Structure, Secondary; D051844:alpha-Synuclein / Q000737:chemistry* / Q000633:toxicity*
chemical_list: D003432:Cross-Linking Reagents; D018696:Neuroprotective Agents; D051844:alpha-Synuclein; D006493:Heparin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Cell Permeabilization; GAPDH; Molecular Modeling; Parkinson Disease; Protofibril Structure; X-ray Scattering; α-Synuclein