Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding

Parasitic organisms especially those of the Apicomplexan phylum, harbour a cytosol localised canonical Hsp70 chaperone. One of the defining features of this protein is the presence of GGMP repeat residues sandwiched between α-helical lid and C-terminal EEVD motif. The role of the GGMP repeats of Hsp70s remains unknown. In the current study, we introduced GGMP mutations in the cytosol localised Hsp70-1 of Plasmodium falciparum (PfHsp70-1) and a chimeric protein (KPf), constituted by the ATPase domain of E. coli DnaK fused to the C-terminal substrate binding domain of PfHsp70-1. A complementation assay conducted using E. coli dnaK756 cells demonstrated that the GGMP motif was essential for chaperone function of the chimeric protein, KPf. Interestingly, insertion of GGMP motif of PfHsp70-1 into DnaK led to a lethal phenotype in E. coli dnaK756 cells exposed to elevated growth temperature. Using biochemical and biophysical assays, we established that the GGMP motif accounts for the elevated basal ATPase activity of PfHsp70-1. Furthermore, we demonstrated that this motif is important for interaction of the chaperone with peptide substrate and a co-chaperone, PfHop. Our findings suggest that the GGMP may account for both the specialised chaperone function and reportedly high catalytic efficiency of PfHsp70-1.

Publication year: 2021
Authors: S. Makumire 1,2 , T. Harmfree Dongola 1, G. Chakafana 1,3, L. Tshikonwane 1, C. Tshikani Chauke 1, T. Maharaj 4, T. Zininga 1,4, A. Shonhai 1
1 – Department of Biochemistry, University of Venda, Private Bag X5050, Thohoyandou 0950, South Africa
2 – Structural Biology Research Unit, Department of Integrative Biomedical Sciences, University of Cape Town, Observatory 7925, South Africa
3 – Department of Medicine, University of Cape Town, Faculty of Health Sciences, Observatory, Cape Town 7925, South Africa
4 – Department of Biochemistry, Stellenbosch University, Stellenbosch 7600, South Africa
Published in: International Journal of Molecular Sciences, 2021, Vol. 22(4), p. 2226
DOI: 10.3390/ijms22042226


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