Banca de DEFESA: MATEUS FARIAS DE SOUZA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : MATEUS FARIAS DE SOUZA
DATE: 23/02/2024
TIME: 14:15
LOCAL: Sala Virtual Remora
TITLE:

Trypanothione reductase of Leishmania braziliensis as a molecular target for the prospecting of new inhibitors: In vitro and in silico evaluation of Gyroxin and Convulxin from Crotalus durissus terrificus as inhibitors

KEY WORDS:

Trypanothione reductase. Leishmania braziliensis. Snake toxins. Convulxin. Gyroxin.

PAGES: 90
BIG AREA: Ciências Biológicas
AREA: Biologia Geral
SUMMARY:

The Trypanothione Reductase (TR) is a validated molecular target for the development of new inhibitors to treat diseases caused by Trypanosomatids. On another front, snake venom toxins, due to their structural and functional diversity, are important sources of bioactive molecules for the development of new drugs, including the treatment of diseases such as Leishmaniasis. In this context, the present study aims to analyze the interaction of toxins (Giroxin and Convulxin from Crotalus durissus terrificus) with the Trypanothione Reductase of L. braziliensis (TRLb) through in silico models and evaluate their inhibition through in vitro models. To achieve this, molecular docking between TRLb and toxins was performed using the ClusPro2.0 tool, and molecular dynamics were conducted using the GROMACS tool. Subsequently, the gene for TRLb was inserted into the pET 28(a+) vector and expressed in E. coli. The relative molecular mass and isoelectric point were determined by 12.5% one-dimensional and two-dimensional SDS-PAGE, and enzymatic activity was evaluated using colorimetric methods. The venom from Crotalus durissus terrificus was obtained from the LABIOPROT-FIOCRUZ-RO venom bank. It was fractionated by affinity chromatography on a Tricorn 10/20 column filled with Benzamidine-Sepharose resin (GE Healthcare). In the inhibition assays, the IC50 was calculated using the non-linear regression model in the GraphPad Prism program. As results, in silico assays indicated that Convulxin and Giroxin are capable of interacting with the enzyme's catalytic site, opening possibilities for further research in the field. TRLb was obtained with a high degree of purity, and its catalytic activity was preserved, with a molecular mass and isoelectric point consistent with those described in the literature. The purification of Giroxin from C.d.t resulted in three fractions, named 1, 2, and Giroxin, with electrophoretic profiling showing a single protein band with a molecular mass of approximately 30 kDa. Giroxin was used at different concentrations for the assessment of its inhibitory capacity, namely 5 μM, 1 μM, and 0.2 μM, resulting in a decrease in the catalytic activity of TRLb by 88%, 41.2%, and 18%, respectively.

COMMITTEE MEMBERS:
Externo à Instituição - ALDO RODRIGUES DA SILVA - UFF
Presidente - ***.161.386-** - ANDREIMAR MARTINS SOARES - FIOCRUZ
Externo à Instituição - LUIS ANDRE MORAIS MARIÚBA - FIOCRUZ