title: Structural and Thermodynamic Properties of Septin 3 Investigated by Small-Angle X-Ray Scattering.
abstract: Septins comprise a family of proteins involved in a variety of cellular processes and related to several human pathologies. They are constituted by three structural domains: the N- and C-terminal domains, highly variable in length and composition, and the central domain, involved in the guanine nucleotide (GTP) binding. Thirteen different human septins are known to form heterogeneous complexes or homofilaments, which are stabilized by specific interactions between the different interfaces present in the domains. In this work, we have investigated by in-solution small-angle x-ray scattering the structural and thermodynamic properties of a human septin 3 construct, SEPT3-GC, which contains both of both interfaces (G and NC) responsible for septin-septin interactions. In order to shed light on the role of these interactions, small-angle x-ray scattering measurements were performed in a wide range of temperatures, from 2 up to 56°C, both with and without a nonhydrolysable form of GTP (GTPγS). The acquired data show a temperature-dependent coexistence of monomers, dimers, and higher-order aggregates that were analyzed using a global fitting approach, taking into account the crystallographic structure of the recently reported SEPT3 dimer, PDB:3SOP. As a result, the enthalpy, entropy, and heat capacity variations that control the dimer-monomer dissociation equilibrium in solution were derived and GTPγS was detected to increase the enthalpic stability of the dimeric species. Moreover, a temperature increase was observed to induce dissociation of SEPT3-GC dimers into monomers just preceding their reassembling into amyloid aggregates, as revealed by the Thioflavin-T fluorescence assays.
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente and Consorzio Nazionale Interuniversitario per le Scienze Fisiche della Materia, Università Politecnica delle Marche, Ancona, Italy.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Ortore
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, Brazil.
authors.forename: Joci N A
authors.initials: JN
authors.lastname: Macedo
authors.affiliation: Instituto de Física de São Carlos, Universidade de São Paulo, São Carlos, Brazil.
authors.forename: Ana Paula U
authors.initials: AP
authors.lastname: Araujo
authors.affiliation: European Synchrotron Radiation Facility, Grenoble, France.
authors.forename: Claudio
authors.initials: C
authors.lastname: Ferrero
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente and Consorzio Nazionale Interuniversitario per le Scienze Fisiche della Materia, Università Politecnica delle Marche, Ancona, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Mariani
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente and Consorzio Nazionale Interuniversitario per le Scienze Fisiche della Materia, Università Politecnica delle Marche, Ancona, Italy. Electronic address: f.spinozzi@univpm.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Spinozzi
authors.affiliation: Instituto de Física da Universidade de São Paulo, São Paulo, Brazil. Electronic address: itri@if.usp.br.
authors.forename: Rosangela
authors.initials: R
authors.lastname: Itri
mesh_terms: D000595:Amino Acid Sequence; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D053838:Scattering, Small Angle; D058112:Septins / Q000737:chemistry* / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D006160:Guanosine Triphosphate; D058112:Septins; C407343:septin 3