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

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dc.contributor.authorMakumire, Stanley
dc.contributor.authorDongola, Tendamudzimu Harmfree
dc.contributor.authorChakafana, Graham
dc.contributor.authorTshikonwane, Lufuno
dc.contributor.authorChauke, Cecilia Tshikani
dc.contributor.authorMaharaj, Tarushai
dc.contributor.authorZininga, Tawanda
dc.contributor.authorShonhai, Addmore
dc.date.accessioned2021-10-19T15:43:36Z
dc.date.available2021-10-19T15:43:36Z
dc.date.issued2021-02-23
dc.date.updated2021-02-26T14:57:18Z
dc.description.abstractParasitic 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.en_US
dc.identifierdoi: 10.3390/ijms22042226
dc.identifier.apacitationMakumire, S., Dongola, T. H., Chakafana, G., Tshikonwane, L., Chauke, C. T., Maharaj, T., ... Shonhai, A. (2021). Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding. <i>International Journal of Molecular Sciences</i>, 22(4), http://hdl.handle.net/11427/35275en_ZA
dc.identifier.chicagocitationMakumire, Stanley, Tendamudzimu Harmfree Dongola, Graham Chakafana, Lufuno Tshikonwane, Cecilia Tshikani Chauke, Tarushai Maharaj, Tawanda Zininga, and Addmore Shonhai "Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding." <i>International Journal of Molecular Sciences</i> 22, 4. (2021) http://hdl.handle.net/11427/35275en_ZA
dc.identifier.citationMakumire, S., Dongola, T.H., Chakafana, G., Tshikonwane, L., Chauke, C.T., Maharaj, T., Zininga, T. & Shonhai, A. et al. 2021. Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding. <i>International Journal of Molecular Sciences.</i> 22(4) http://hdl.handle.net/11427/35275en_ZA
dc.identifier.ris TY - Journal Article AU - Makumire, Stanley AU - Dongola, Tendamudzimu Harmfree AU - Chakafana, Graham AU - Tshikonwane, Lufuno AU - Chauke, Cecilia Tshikani AU - Maharaj, Tarushai AU - Zininga, Tawanda AU - Shonhai, Addmore AB - 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. DA - 2021-02-23 DB - OpenUCT DP - University of Cape Town IS - 4 J1 - International Journal of Molecular Sciences KW - malaria KW - Plasmodium falciparum KW - chaperone KW - GGMP repeats KW - Hsp70 KW - Hop LK - https://open.uct.ac.za PY - 2021 T1 - Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding TI - Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding UR - http://hdl.handle.net/11427/35275 ER - en_ZA
dc.identifier.urihttps://doi.org/10.3390/ijms22042226
dc.identifier.urihttp://hdl.handle.net/11427/35275
dc.identifier.vancouvercitationMakumire S, Dongola TH, Chakafana G, Tshikonwane L, Chauke CT, Maharaj T, et al. Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding. International Journal of Molecular Sciences. 2021;22(4) http://hdl.handle.net/11427/35275.en_ZA
dc.language.isoenen_US
dc.publisher.departmentDepartment of Integrative Biomedical Sciencesen_US
dc.publisher.facultyFaculty of Health Sciencesen_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.sourceInternational Journal of Molecular Sciencesen_US
dc.source.journalissue4en_US
dc.source.journalvolume22en_US
dc.source.urihttps://www.mdpi.com/journal/ijms
dc.subjectmalaria
dc.subjectPlasmodium falciparum
dc.subjectchaperone
dc.subjectGGMP repeats
dc.subjectHsp70
dc.subjectHop
dc.titleMutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Bindingen_US
dc.typeJournal Articleen_US
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