title: Protein thermal denaturation is modulated by central residues in the protein structure network.
abstract: Network structural analysis, known as residue interaction networks or graphs (RIN or RIG, respectively) or protein structural networks or graphs (PSN or PSG, respectively), comprises a useful tool for detecting important residues for protein function, stability, folding and allostery. In RIN, the tertiary structure is represented by a network in which residues (nodes) are connected by interactions (edges). Such structural networks have consistently presented a few central residues that are important for shortening the pathways linking any two residues in a protein structure. To experimentally demonstrate that central residues effectively participate in protein properties, mutations were directed to seven central residues of the β-glucosidase Sfβgly (β-D-glucoside glucohydrolase; EC 3.2.1.21). These mutations reduced the thermal stability of the enzyme, as evaluated by changes in transition temperature (Tm ) and the denaturation rate at 45 °C. Moreover, mutations directed to the vicinity of a central residue also caused significant decreases in the Tm of Sfβgly and clearly increased the unfolding rate constant at 45 °C. However, mutations at noncentral residues or at surrounding residues did not affect the thermal stability of Sfβgly. Therefore, the data reported in the present study suggest that the perturbation of the central residues reduced the stability of the native structure of Sfβgly. These results are in agreement with previous findings showing that networks are robust, whereas attacks on central nodes cause network failure. Finally, the present study demonstrates that central residues underlie the functional properties of proteins.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Brazil.
authors.forename: Valquiria P
authors.identifier: http://orcid.org/0000-0002-3052-2112
authors.initials: VP
authors.lastname: Souza
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Brazil.
authors.forename: Cecília M
authors.initials: CM
authors.lastname: Ikegami
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Brazil.
authors.forename: Guilherme M
authors.initials: GM
authors.lastname: Arantes
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Brazil.
authors.forename: Sandro R
authors.initials: SR
authors.lastname: Marana
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002942:Circular Dichroism; D004795:Enzyme Stability; D006358:Hot Temperature; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D060066:Protein Interaction Maps; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D018411:Spodoptera / Q000201:enzymology / Q000235:genetics; D001617:beta-Glucosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D011506:Proteins; D001617:beta-Glucosidase
keywords: Spodoptera frugiperda; glycoside hydrolase; protein structural network; residue interaction network; β-glucosidase