Recombinant Expression and Biophysical Characterization of a Druggable Schistosoma mansoni Universal Stress G4LZI3 Protein

Abiola Fatimah Adenowo; Priscilla Masamba; Ndibonani Kebonang Qokoyi; Babatunji Emmanuel Oyinloye; Abidemi Paul Kappo

doi:10.34172/apb.2022.035

Recombinant Expression and Biophysical Characterization of a Druggable Schistosoma mansoni Universal Stress G4LZI3 Protein

Adv Pharm Bull. 2022 Mar;12(2):366-374. doi: 10.34172/apb.2022.035. Epub 2021 Jan 31.

Authors

Abiola Fatimah Adenowo¹, Priscilla Masamba², Ndibonani Kebonang Qokoyi², Babatunji Emmanuel Oyinloye³, Abidemi Paul Kappo²

Affiliations

¹ Department of Medical Biochemistry, Faculty of Basic Medical Sciences, Lagos State University College of Medicine, PMB 21266, Ikeja, Lagos, Nigeria.
² Molecular Biophysics and Structural Biology (MBSB) Group, Department of Biochemistry, University of Johannesburg, Kingsway Campus, Auckland Park 2006, South Africa.
³ Department of Biochemistry, Afe Babalola University, PMB 5454, Ado-Ekiti 360001, Nigeria.

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.

Keywords: Biophysical characterization; G4LZI3; Recombinant proteins; Schistosoma mansoni; Schistosomiasis.