A Cell Model for HSP60 Deficiencies: Modeling Different Levels of Chaperonopathies Leading to Oxidative Stress and Mitochondrial Dysfunction

  • Cagla Cömert
  • Paula Fernandez-Guerra
  • Peter BrossEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1873)


Besides providing the majority of ATP production in cells, mitochondria are also involved in many other cellular functions and are central for cellular stress signaling. Mitochondrial dysfunction induces not only inherited mitochondrial disorders but also contributes to neurodegenerative diseases, cancer, diabetes, and metabolic syndrome. The HSP60/HSP10 molecular chaperone complex facilitates folding of mitochondrial proteins and is thus an important factor for many mitochondrial functions. To model different degrees of oxidative stress and mitochondrial dysfunction we here describe a HEK293 derived Flp-In cell system with stable insertion and tunable expression of HSP60 cDNA carrying a dominant negative mutation. When expressed the dominant negative HSP60 mutant is incorporated into endogenously encoded HSP60/HSP10 complexes and impairs chaperone activity of the HSP60/HSP10 complex in a dose dependent manner. Using this system, different levels of oxidative stress and mitochondrial dysfunction challenges can be generated depending on the induction level of the mutant HSP60 cDNA insert. Here we describe our system and pertinent analysis methodology for use in studies of mitochondrial chaperone deficiency and resulting effects of increased production of reactive oxygen species and mitochondrial dysfunction.


Heat shock protein 60 deficiency Cell models Image cytometry Mass spectrometry 



We acknowledge financial support from the Ludvig and Sara Elsass Foundation and the Eva og Henry Frænkels Mindefond. C.C. was supported by a scholarship from SCITECH, Aarhus University. We thank Anne Bie for introduction to the Flp-in cell model.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Cagla Cömert
    • 1
    • 2
  • Paula Fernandez-Guerra
    • 1
    • 2
  • Peter Bross
    • 1
    • 2
    Email author
  1. 1.Research Unit for Molecular Medicine, Department of Clinical Medicine, HEALTHAarhus University, and Department of Clinical Biochemistry Aarhus University HospitalAarhusDenmark
  2. 2.Department of BiochemistryAarhus University HospitalAarhusDenmark

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