Banca de DEFESA: Gabriel dos Santos Silva

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : Gabriel dos Santos Silva
DATE: 06/10/2023
TIME: 14:00
LOCAL: Microsoft Teams
TITLE:

: Integration of Computational and Experimental Approaches to Characterize a Salivary Odorant-Binding Protein from Rhodnius neglectus, Vector of Chagas Disease

KEY WORDS:

Proteína ligante de odorante; Rhodnius neglectus; Dinâmica molecular; Biologia estrutural.

PAGES: 65
BIG AREA: Ciências da Saúde
AREA: Medicina
SUMMARY:

Rhodnius neglectus, an insect from the Reduviidae family (Hemiptera), is a potential vector of Trypanosoma cruzi, the causative protozoan of Chagas disease. Triatomine saliva comprises a rich mixture of molecules synthesized by the salivary glands to counter host responses, aid the uptake of blood and, indirectly, T. cruzi transmission. Therefore, the saliva of this hematophagous insect has been a relevant subject of studies due to the presence of these molecules with high biotechnological potential. The goal of this work was to study an Odorant Binding Protein from R. neglectus saliva (RnOBP) using computational and experimental approaches aiming, to contribute to the understanding of its structural and functional characteristics. RnOBP molecular modeling, computational molecular dynamics and clustering tests were performed to obtain information about the protein's flexibility. Next, molecular docking assays were carried out to verify the affinity of RnOBP with agonists of vertebrate coagulation pathways. Finally, putative RnOBP oligomers were observed. To obtain the recombinant RnOBP, its nucleotide sequence was obtained from the R. neglectus transcriptome, cloned into the pET100/D-TOPO expression vector and produced by Escherichia coli Rosetta(DE3)pLysS bacteria. The recombinant RnOBP was purified by affinity chromatography, refolded, and used in biophysical characterization assays. This protein is modeled to be entirely formed by α-helices and have a significant internal cavity, potentially a ligand-binding site. Compared with insect OBPs, regardless of the composition or classification of the OBP, the binding site region is well conserved. The recombinant RnOBP purified from inclusion bodies revealed a secondary structure composition different than expected. Initial functional indicated rRnOBP could act to inhibit platelet aggregation, but not coagulation. This study showed RnOBP structural properties and may contribute to the understanding of its functions and biotechnological potential. 

COMMITTEE MEMBERS:
Presidente - 2088007 - CARLA NUNES DE ARAUJO
Externo à Instituição - CLEVER GOMES CARDOSO
Externa ao Programa - ***.924.081-** - PAULA BEATRIZ DE MEDEIROS SANTIAGO - UnB