Affinity purification probes of potential use to investigate the endogenous Hsp70 interactome in cancer.

TitleAffinity purification probes of potential use to investigate the endogenous Hsp70 interactome in cancer.
Publication TypeJournal Article
Year of Publication2014
AuthorsRodina 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
JournalACS Chem Biol
Volume9
Issue8
Pagination1698-705
Date Published2014 Aug 15
ISSN1554-8937
KeywordsAffinity 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.

DOI10.1021/cb500256u
Alternate JournalACS Chem. Biol.
PubMed ID24934503
PubMed Central IDPMC4134716
Grant ListCA158728 / 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