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Purification and biochemical characterization of Msi3, an essential Hsp110 molecular chaperone in Candida albicans

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Cell Stress and Chaperones Aims and scope

Abstract

Hsp110s are unique and essential molecular chaperones in the eukaryotic cytosol. They play important roles in maintaining cellular protein homeostasis. Candida albicans is the most prevalent yeast opportunistic pathogen that causes fungal infections in humans. As the only Hsp110 in Candida albicans, Msi3 is essential for the growth and infection of Candida albicans. In this study, we have expressed and purified Msi3 in nucleotide-free state and carried out biochemical analyses. Sse1 is the major Hsp110 in budding yeast S. cerevisiae and the best characterized Hsp110. Msi3 can substitute Sse1 in complementing the temperature-sensitive phenotype of S. cerevisiae carrying a deletion of SSE1 gene although Msi3 shares only 63.4% sequence identity with Sse1. Consistent with this functional similarity, the purified Msi3 protein shares many similar biochemical activities with Sse1 including binding ATP with high affinity, changing conformation upon ATP binding, stimulating the nucleotide-exchange for Hsp70, preventing protein aggregation, and assisting Hsp70 in refolding denatured luciferase. These biochemical characterizations suggested that Msi3 can be used as a model for studying the molecular mechanisms of Hsp110s.

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Acknowledgements

We thank Dr. Ronda J. Rolfes (Department of Biology, Georgetown University) for providing the genomic DNA of C. albicans. We thank Ms. Baoxiu Zhang for technical support and Mr. Liqing Hu and Justin Kidd for discussion.

Funding

This work was supported by NIH (R01GM098592 and R21AI140006 to Q.L.), and VETAR Award from VCU (to Q.L.).

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  1. Ying Wang, Hongtao Li and Cancan Sun contributed equally to this work.

Authors and Affiliations

  1. Department of Physiology and Biophysics, Virginia Commonwealth University, Richmond, VA, 23298, USA

    Ying Wang, Hongtao Li, Cancan Sun, Lei Zhou & Qinglian Liu

  2. Key Laboratory of Food Nutrition and Safety, Tianjin University of Science & Technology, Tianjin, 300457, China

    Qingdai Liu

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  1. Ying Wang
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  2. Hongtao Li
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Correspondence to Qinglian Liu.

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Wang, Y., Li, H., Sun, C. et al. Purification and biochemical characterization of Msi3, an essential Hsp110 molecular chaperone in Candida albicans. Cell Stress and Chaperones 26, 695–704 (2021). https://doi.org/10.1007/s12192-021-01213-5

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