Abstract
Secreted, or extracellular, heat shock protein 90 (eHsp90) is considered a recent discovery in eukaryotes. Over the last two decades, studies have provided significant supporting evidence that implicates eHsp90 both in normal cellular processes such as wound healing and in the development of human pathologies and diseases including fibrosis and cancer. In the early 2000s, Eustace et al. demonstrated that eHsp90 promotes the invasion of breast cancer cells by binding to and regulating the activity of an extracellular matrix (ECM) remodeling enzyme, the matrix metalloproteinase 2 or MMP2. Interestingly, inside mammalian cells, Hsp90 is an essential chaperone that interacts with hundreds of newly synthesized proteins, known as “clients,” that require Hsp90’s assistance to perform their function. Several methods are routinely used to characterize the role and impact of Hsp90 on a client protein’s functionality in vitro and in vivo. However, the mechanistic role of eHsp90 is less well-defined since, so far, only a handful of extracellular client proteins have been identified. Here, we describe methods to characterize the impact of the secreted chaperone on MMP2 activity, the most characterized extracellular client of eHsp90. The procedures described here can be applied and adapted to characterize other extracellular clients, particularly members of the MMP family.
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Acknowledgments
We would like to thank Dr. Alexander J. Baker-Williams for critical discussions and technical support. Dimitra Bourboulia is supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number R01GM139932. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Additional funding sources include the SUNY Upstate Medical University, the Upstate Foundation, the SUNY Research Foundation, and the Carol M. Baldwin Breast Cancer Research fund grant (D.B.).
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Votra, S.D., Alsalih, D., Bourboulia, D. (2023). Methods to Assess the Impact of Hsp90 Chaperone Function on Extracellular Client MMP2 Activity. In: Calderwood, S.K., Prince, T.L. (eds) Chaperones. Methods in Molecular Biology, vol 2693. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3342-7_17
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DOI: https://doi.org/10.1007/978-1-0716-3342-7_17
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