Abstract
Purpose: Universal stress protein (USP) from Schistosoma mansoni, designated as G4LZI3, waspreviously hypothesised as a druggable target and vaccine candidate for human schistosomiasis.The purpose of this study is to characterize a purified recombinant G4LZI3 preliminarily forsubsequent structural characterization, which will provide baseline structural data for futurefunctional studies for the discovery, design and development of new schistosomal drugs for thetreatment, control and elimination of schistosomiasis. Methods: Restriction digest analysis of a GenScript-synthesised codon-optimised G4LZI3gene construct was carried out to ascertain its integrity and size. Thereafter, the pQE30-G4LZI3 construct was transformed into an M15 bacterial expression host. Transformed cellswere induced with isopropyl β-D-thiogalactoside for recombinant protein expression of anappreciable amount of pQE30-G4LZI3, which was subsequently purified with fast proteinliquid chromatography (FPLC) and a size exclusion chromatographic purification scheme.Preliminary biophysical characterization of the 6X His-tagged G4LZI3 was done to determineits secondary structure characteristics and protein stability. Results: A molecular weight protein of 20.3 kDa was confirmed subsequent to restriction digestanalysis, while heterologous protein expression yielded a highly soluble and considerableamount of histidine-tagged G4LZI3 protein, which was successfully purified to homogeneity.Biophysical characterization indicated that the protein was well folded, heat-stable, had thefunctional groups and secondary structure composition required and was thus amenable tofurther structural characterization and determination. Conclusion: Biophysical characterization of purified G4LZI3 showed that further structuralstudies can be embarked upon on the use of G4LZI3 as a druggable target and possibly avaccine target against schistosomiasis via vaccinomics.
Highlights
Human schistosomiasis persists as one of the neglected tropical diseases, with over 261 million individuals infected worldwide; a great proportion of these are located in Africa, South America and Asia
Transformed cells were induced with isopropyl β-D-thiogalactoside for recombinant protein expression of an appreciable amount of pQE30-G4LZI3, which was subsequently purified cwith fast protein liquid chromatography and a size exclusion chromatographic purification scheme
Biophysical characterization of purified G4LZI3 showed that further structural studies can be embarked upon on the use of G4LZI3 as a druggable target and possibly a vaccine target against schistosomiasis via vaccinomics
Summary
Human schistosomiasis persists as one of the neglected tropical diseases, with over 261 million individuals infected worldwide; a great proportion of these are located in Africa, South America and Asia. It is second to malaria in terms of prevalence and is the second top neglected tropical disease after hookworm infection in subt Saharan Africa.[1,2] This debilitating parasitic disease is caused by the trematode-worms of the genus Schistosoma Three members of this class are known to infect humans: Schistosoma mansoni and Schistosoma japonicum are responsible for intestinal schistosomiasis, while Schistosoma haematobium causes urinary schistosomiasis.[2,3] ip The lifecycle of the blood-dwelling flatworms involves two developmental phases, a sexual phase in the definitive human host and an asexual phase in the freshwater snail host.[4,5] Schistosomiasis occurs owing to r immunological reactions caused by the Schistosoma eggs entrapped in tissues. These include health education and improved sanitation, use of biological control approaches to eliminate the u intermediate snail host of the parasites and mass administration of Praziquantel (PZQ).[2,9] So far, PZQ, a pyrazinoisoquinole antihelminthic, is the only frontline drug for the treatment of all forms of schistomiasis
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