In silico analysis and molecular identification of an anaphase-promoting complex homologue from human pathogen Entamoeba histolytica

• Published in: Journal of Genetic Engineering and Biotechnology Vol. 19; no. 1; pp. 133 - 10
• Main Authors: Pal, Suchetana, Biswas, Pinaki, Ghosh, Raktim, Dam, Somasri
• Format: Journal Article
• Language: English
• Published: Cairo Springer 01-09-2021; Springer Nature B.V; Springer Berlin Heidelberg; Elsevier
• Subjects: Analysis; Anaphase; Anaphase-promoting complex; Antibiotic resistance; Antibiotics; Antigenicity; APC; Automation; Binding sites; Bioinformatics; Carcinogens; Care and treatment; Cell cycle; Cell division; Chromosomes; Conserved sequence; Developing countries; Drug dosages; Drug resistance; Drug resistance in microorganisms; E coli; Entamoeba histolytica; Enzymes; Genetic aspects; Glutathione; Glutathione transferase; Health aspects; Homology; Homology modelling; In silico analysis; India; LDCs; Ligands; Ligases; Localization; Malaria; Metronidazole; Molecular identification; Molecular weight; Morbidity; Mortality; Parasites; Pathogens; Phylogenetics; Proteasomes; Protein interaction; Protein-protein interactions; Proteins; Structure-function relationships; Substrates; Therapeutic targets; Three dimensional models; Ubiquitin; Ubiquitin-protein ligase; India
• ISSN: 1687-157X; 2090-5920
• DOI: 10.1186/s43141-021-00234-y

Background Amoebiasis, being endemic worldwide, is the second leading cause of parasite-associated morbidity and mortality after malaria. The human parasite Entamoeba histolytica is responsible for the disease. Metronidazole is considered as the gold standard for the treatment of amoebiasis, but this antibiotic is carcinogenic and the development of antibiotic resistance against E. histolytica is a major health concern. Chromosome segregation is irregular in this parasite due to the absence of a few cell cycle checkpoint proteins. Anaphase-promoting complex (APC/C or cyclosome) is an E3 ubiquitin ligase that synchronizes chromosome segregation and anaphase progression via the ubiquitin-proteasome system. Proteasome is considered to be an attractive drug target for protozoan parasites. For the present study, EhApc11 from E. histolytica, a homologue of Apc11 in humans, is selected for elucidating its structural and functional aspects by detailed in silico analysis and molecular methods. Its physicochemical characteristics, identification of probable interactors, 3D model and quality analysis are done using standard bioinformatics tools. cDNA sequence of EhAPC11 has been further cloned for molecular characterization. Result Conserved domain analysis revealed that EhApc11 belongs to the RING (really interesting new gene) superfamily and has ligand binding capacity. Expression study in Escherichia coli BL21 (DE3) revealed that the molecular weight of glutathione S-transferase (GST)-tagged protein is ~ 36 kDa. Conclusion EhApc11 is a hydrophilic, thermostable, extracellular protein with potent antigenicity. The study will serve as a groundwork for future in-depth analysis regarding the validation of protein-protein interaction of EhApc11 with its substrates identified by STRING analysis and the potential of EhApc11 to serve as an anti-amoebic drug target.