Title | Affinity purification probes of potential use to investigate the endogenous Hsp70 interactome in cancer. |
Publication Type | Journal Article |
Year of Publication | 2014 |
Authors | Rodina A, Taldone T, Kang Y, Patel PD, Koren J, Yan P, Gomes EMDaGama, Yang C, Patel MR, Shrestha L, Ochiana SO, Santarossa C, Maharaj R, Gozman A, Cox MB, Erdjument-Bromage H, Hendrickson RC, Cerchietti L, Melnick A, Guzman ML, Chiosis G |
Journal | ACS Chem Biol |
Volume | 9 |
Issue | 8 |
Pagination | 1698-705 |
Date Published | 2014 Aug 15 |
ISSN | 1554-8937 |
Keywords | Affinity Labels, Cell Line, Tumor, Chromatography, Affinity, HSP70 Heat-Shock Proteins, Humans, Neoplasms, Protein Binding |
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. |
DOI | 10.1021/cb500256u |
Alternate Journal | ACS Chem. Biol. |
PubMed ID | 24934503 |
PubMed Central ID | PMC4134716 |
Grant List | CA158728 / CA / NCI NIH HHS / United States P30 CA008748 / CA / NCI NIH HHS / United States P30 CA08748 / CA / NCI NIH HHS / United States R01 CA119001 / CA / NCI NIH HHS / United States R01 CA119001 / CA / NCI NIH HHS / United States R01 CA155226 / CA / NCI NIH HHS / United States R01 CA155226 / CA / NCI NIH HHS / United States R01 CA172546 / CA / NCI NIH HHS / United States R01 CA172546 / CA / NCI NIH HHS / United States R21 CA158609 / CA / NCI NIH HHS / United States UL1 RR024996 / RR / NCRR NIH HHS / United States |