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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 Hsp...

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Autores principales: Makumire, Stanley, Dongola, Tendamudzimu Harmfree, Chakafana, Graham, Tshikonwane, Lufuno, Chauke, Cecilia Tshikani, Maharaj, Tarushai, Zininga, Tawanda, Shonhai, Addmore
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926355/
https://www.ncbi.nlm.nih.gov/pubmed/33672387
http://dx.doi.org/10.3390/ijms22042226
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author Makumire, Stanley
Dongola, Tendamudzimu Harmfree
Chakafana, Graham
Tshikonwane, Lufuno
Chauke, Cecilia Tshikani
Maharaj, Tarushai
Zininga, Tawanda
Shonhai, Addmore
author_facet Makumire, Stanley
Dongola, Tendamudzimu Harmfree
Chakafana, Graham
Tshikonwane, Lufuno
Chauke, Cecilia Tshikani
Maharaj, Tarushai
Zininga, Tawanda
Shonhai, Addmore
author_sort Makumire, Stanley
collection PubMed
description 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.
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spelling pubmed-79263552021-03-04 Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding Makumire, Stanley Dongola, Tendamudzimu Harmfree Chakafana, Graham Tshikonwane, Lufuno Chauke, Cecilia Tshikani Maharaj, Tarushai Zininga, Tawanda Shonhai, Addmore Int J Mol Sci Article 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. MDPI 2021-02-23 /pmc/articles/PMC7926355/ /pubmed/33672387 http://dx.doi.org/10.3390/ijms22042226 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Makumire, Stanley
Dongola, Tendamudzimu Harmfree
Chakafana, Graham
Tshikonwane, Lufuno
Chauke, Cecilia Tshikani
Maharaj, Tarushai
Zininga, Tawanda
Shonhai, Addmore
Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding
title Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding
title_full Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding
title_fullStr Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding
title_full_unstemmed Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding
title_short Mutation of GGMP Repeat Segments of Plasmodium falciparum Hsp70-1 Compromises Chaperone Function and Hop Co-Chaperone Binding
title_sort mutation of ggmp repeat segments of plasmodium falciparum hsp70-1 compromises chaperone function and hop co-chaperone binding
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926355/
https://www.ncbi.nlm.nih.gov/pubmed/33672387
http://dx.doi.org/10.3390/ijms22042226
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