Abstract
Heat
shock protein 70 (Hsp70) is a family of proteins with key roles in
regulating malignancy. Cancer cells rely on Hsp70 to inhibit apoptosis,
regulate senescence and autophagy, and maintain the stability of numerous
onco-proteins. Despite these important biological functions in cancer,
robust chemical tools that enable the analysis of the Hsp70-regulated
proteome in a tumor-by-tumor manner are yet unavailable. Here we take
advantage of a recently reported Hsp70 ligand to design and develop
an affinity purification chemical toolset for potential use in the
investigation of the endogenous Hsp70-interacting proteome in cancer.
We demonstrate that these tools lock Hsp70 in complex with onco-client
proteins and effectively isolate Hsp70 complexes for identification
through biochemical techniques. Using these tools we provide proof-of-concept
analyses that glimpse into the complex roles played by Hsp70 in maintaining
a multitude of cell-specific malignancy-driving proteins.