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Characterization And Structure‐Function Analysis of Autophagy Factor ATG7 from Trypanosoma Brucei (TbATG7) as a Potential Drug Target
Author(s) -
Karnes Kayla,
Shaikh Khalid
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.03861
Subject(s) - trypanosoma brucei , autophagy , in silico , biology , african trypanosomiasis , gene , computational biology , genetics , microbiology and biotechnology , trypanosomiasis , virology , apoptosis
The characterization of autophagy pathways is a relatively new area of study, but it is one that shows great promise for research, especially in kinetoplastids. This classification of organisms includes Trypanosoma brucei (T. brucei) , the causative agent of Human African Trypanosomiasis, otherwise known as African Sleeping Sickness. This disease is often fatal, and its current diagnosis and treatment regimens are complex and ineffective, so there is a requirement for a new potential drug target. Autophagy has been shown to be important for the survival of kinetoplastids during starvation and differentiation, which suggests that the autophagy pathway could be a target for treatment of T. brucei infections. In this study, the autophagy‐related gene 7 in T. brucei (TbATG7) is characterized and analyzed using heterologous expression for protein and in silico . Overall, this research focuses on the hypothesis that the characterization of TbATG7 with the inhibitors directed against recombinant protein could inhibit the growth of the parasite, thus making it a potential drug target. The TbATG7 gene was successfully amplified using touch‐down polymerase chain reaction. Furthermore, the plasmid which will be used to make a construct, pUC19, transformed into BL‐21 competent cells, was successfully cleaved using respective restriction enzymes. The bioinformatic analysis of this research can be classified into two major aims. The first aim was to analyze the TbATG7 protein from sequence to structure, including sequence homology, homology modeling, and a three‐dimensional (3D) model for structural analysis. The second aim would be to determine binding sites to control protein function based on the structural analysis. The preliminary results has provided a predicted 3D protein structure model using SWISS‐Model and PyMol, as well as multiple sequence alignment of the protein's sequence in comparison to other related organisms protein sequences. The active site triad, ATP‐binding site and conserved domain were identified on TbATG7 and it shows that TbATG7 belongs to ubiquitin E1‐like protein activating enzyme Apg7 superfamily of proteins. Therefore, the aim of this research is to focus on computational analysis of the TbATG7 protein that will allow to relate protein structure‐function relationship and furthermore to search for targeted screening of inhibitors.