title: Structural analysis of intermolecular interactions in the kinesin adaptor complex fasciculation and elongation protein zeta 1/ short coiled-coil protein (FEZ1/SCOCO).
abstract: Cytoskeleton and protein trafficking processes, including vesicle transport to synapses, are key processes in neuronal differentiation and axon outgrowth. The human protein FEZ1 (fasciculation and elongation protein zeta 1 / UNC-76, in C. elegans), SCOCO (short coiled-coil protein / UNC-69) and kinesins (e.g. kinesin heavy chain / UNC116) are involved in these processes. Exploiting the feature of FEZ1 protein as a bivalent adapter of transport mediated by kinesins and FEZ1 protein interaction with SCOCO (proteins involved in the same path of axonal growth), we investigated the structural aspects of intermolecular interactions involved in this complex formation by NMR (Nuclear Magnetic Resonance), cross-linking coupled with mass spectrometry (MS), SAXS (Small Angle X-ray Scattering) and molecular modelling. The topology of homodimerization was accessed through NMR (Nuclear Magnetic Resonance) studies of the region involved in this process, corresponding to FEZ1 (92-194). Through studies involving the protein in its monomeric configuration (reduced) and dimeric state, we propose that homodimerization occurs with FEZ1 chains oriented in an anti-parallel topology. We demonstrate that the interaction interface of FEZ1 and SCOCO defined by MS and computational modelling is in accordance with that previously demonstrated for UNC-76 and UNC-69. SAXS and literature data support a heterotetrameric complex model. These data provide details about the interaction interfaces probably involved in the transport machinery assembly and open perspectives to understand and interfere in this assembly and its involvement in neuronal differentiation and axon outgrowth.
authors.affiliation: Laboratório Nacional de Biociências-LNBio, Centro Nacional de Pesquisa em Energia e Materiais-CNPEM, Campinas, SP, Brasil.
authors.forename: Marcos Rodrigo
authors.initials: MR
authors.lastname: Alborghetti
authors.forename: Ariane da Silva
authors.initials: Ada S
authors.lastname: Furlan
authors.forename: Júlio César
authors.initials: JC
authors.lastname: da Silva
authors.forename: Maurício Luís
authors.initials: ML
authors.lastname: Sforça
authors.forename: Rodrigo Vargas
authors.initials: RV
authors.lastname: Honorato
authors.forename: Daniela Campos
authors.initials: DC
authors.lastname: Granato
authors.forename: Deivid Lucas
authors.initials: DL
authors.lastname: dos Santos Migueleti
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Neves
authors.forename: Paulo Sergio Lopes
authors.initials: PS
authors.lastname: de Oliveira
authors.forename: Adriana Franco
authors.initials: AF
authors.lastname: Paes-Leme
authors.forename: Ana Carolina de Mattos
authors.initials: AC
authors.lastname: Zeri
authors.forename: Iris Concepcion Linares
authors.initials: IC
authors.lastname: de Torriani
authors.forename: Jörg
authors.initials: J
authors.lastname: Kobarg
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D001692:Biological Transport; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016547:Kinesins; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D053838:Scattering, Small Angle; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques; D014961:X-Ray Diffraction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C528597:FEZ1 protein, human; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; C430910:SCOC protein, human; D016547:Kinesins