Abstract
The HSP60/HSP10 chaperonin assists folding of proteins in the mitochondrial matrix space by enclosing them in its central cavity. The chaperonin forms part of the mitochondrial protein quality control system. It is essential for cellular survival and mutations in its subunits are associated with rare neurological disorders. Here we present the first survey of interactors of the human mitochondrial HSP60/HSP10 chaperonin. Using a protocol involving metabolic labeling of HEK293 cells, cross-linking, and immunoprecipitation of HSP60, we identified 323 interacting proteins. As expected, the vast majority of these proteins are localized to the mitochondrial matrix space. We find that approximately half of the proteins annotated as mitochondrial matrix proteins interact with the HSP60/HSP10 chaperonin. They cover a broad spectrum of functions and metabolic pathways including the mitochondrial protein synthesis apparatus, the respiratory chain, and mitochondrial protein quality control. Many of the genes encoding HSP60 interactors are annotated as disease genes. There is a correlation between relative cellular abundance and relative abundance in the HSP60 immunoprecipitates. Nineteen abundant matrix proteins occupy more than 60% of the HSP60/HSP10 chaperonin capacity. The reported inventory of interactors can form the basis for interrogating which proteins are especially dependent on the chaperonin.
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Funding
We thank the HEALTH graduate school, Aarhus University for support (PhD scholarship and travel and material support to A.B.), and the Hede Nielsens Fond, and the Deutscher Akademischer Austauschdienst (DAAD) for funding of materials, travel and accommodation.
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Anne Bie, Peter Bross, Ulrich Hartl, Roman Körner, Thomas Corydon, Johan Palmfeldt, and Mark Hipp contributed to the study conception and design. Data collection and analysis were performed by Anne Bie, Roman Körner, Johan Palmfeldt, Mark Hipp, Cagla Cömert, and Peter Bross. The first draft of the manuscript was written by Anne Bie and Peter Bross. All authors commented on previous versions of the manuscript, and all authors read and approved the final manuscript. Ulrich Hartl and Peter Bross provided resources and lab-facilities.
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Bie, A.S., Cömert, C., Körner, R. et al. An inventory of interactors of the human HSP60/HSP10 chaperonin in the mitochondrial matrix space. Cell Stress and Chaperones 25, 407–416 (2020). https://doi.org/10.1007/s12192-020-01080-6
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DOI: https://doi.org/10.1007/s12192-020-01080-6