Characterization and biophysical properties of the SIVA1 protein

Author(s)

Larissa Elizabeth Cordeiro Dantas

• Document type: Master's Dissertation
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas
• Defense date: 2015-02-06

Examining board members

Sara Teresinha Olalla Saad; Ljubica Tasic; Ana Olivia Tiroli Cepeda

• Advisor: Sara Teresinha Olalla Saad

Abstract

Siva1 protein interacts with tumor protein p53 and with the member of the tumor necrosis factor receptor superfamily, stathmin, among others. These proteins are related to several pathways involved in cancer and are therefore, strong targets for drug design. Thus, the aim of this study was to characterize the biophysical properties of Siva 1 C-terminal domain in order to contribute to the discovery of new target directed drugs. The C-terminus Siva1 domain (residues 84-175) was fused to gluthatione S-transferase (GST) and expressed in an E coli system and the recombinant GST-Siva C-terminus was purified using GST-Tagged Protein affinity and gel filtration chromatography. We tested the biological activity of the purified Siva C-terminus domain in an extract of the Jurkat cell line that contained natural SIVA binders. Biophysical and biochemical assays showed that the protein was monodisperse in solution with a predominant unfolded and elongated shape. However, at high concentrations, the protein showed a tendency to form soluble aggregates. These results are expected to lead to further progress in better understanding of SIVA1 properties and target-directed drug design. Experiments took place at "Laboratório de Bioquímica de Proteínas" at Institute of Chemistry (IQ-Unicamp), "Laboratório Nacional de Luz Síncrotron" (LNLS) and "Laboratório Nacional de Biociências" (LNBio-LEC) (AU)

• FAPESP's process: 12/07742-0 - Structural characterization and physicochemical properties of SIVA1 protein
• Grantee: Larissa Elizabeth Cordeiro Dantas
• Support Opportunities: Scholarships in Brazil - Master