Structural analysis of intermolecular interactions in the kinesin adaptor complex fasciculation and elongation protein zeta 1/ short coiled-coil protein (FEZ1/SCOCO)

• Published in: PloS one Vol. 8; no. 10; p. e76602
• Main Authors: Alborghetti, Marcos Rodrigo, Furlan, Ariane da Silva, da Silva, Júlio César, Sforça, Maurício Luís, Honorato, Rodrigo Vargas, Granato, Daniela Campos, dos Santos Migueleti, Deivid Lucas, Neves, Jorge L, de Oliveira, Paulo Sergio Lopes, Paes-Leme, Adriana Franco, Zeri, Ana Carolina de Mattos, de Torriani, Iris Concepcion Linares, Kobarg, Jörg
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 07-10-2013; Public Library of Science (PLoS)
• Subjects: Adapters; Adaptor Proteins, Signal Transducing - chemistry; Adaptor Proteins, Signal Transducing - genetics; Adaptor Proteins, Signal Transducing - metabolism; Amino Acid Sequence; Amino acids; Assembly; Binding Sites - genetics; Biochemistry; Biochemistry, Molecular Biology; Biological Transport; Carrier Proteins - chemistry; Carrier Proteins - genetics; Carrier Proteins - metabolism; Chains; Cloning; Coils; Complex formation; Computational neuroscience; Crosslinking; Cytoskeleton; Differentiation; Elongation; Fasciculation; Gene expression; Humans; Interfaces; Kinases; Kinesin; Kinesins; Leukemia; Life Sciences; Magnetic resonance; Magnetic Resonance Spectroscopy; Mass spectrometry; Mass spectroscopy; Membrane Proteins - chemistry; Membrane Proteins - genetics; Membrane Proteins - metabolism; Microtubule-Associated Proteins - metabolism; Models, Molecular; Molecular chains; Molecular modelling; Molecular Sequence Data; Mutation; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; NMR; Nuclear magnetic resonance; Protein Binding; Protein Multimerization; Protein Structure, Tertiary; Protein transport; Proteins; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Resonance; Resonance scattering; Scattering, Small Angle; Sequence Homology, Amino Acid; Small angle X ray scattering; Structural analysis; Structural Biology; Synapses; Topology; Transport; Two-Hybrid System Techniques; X ray scattering; X-Ray Diffraction
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0076602

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.