title: Affinity purification probes of potential use to investigate the endogenous Hsp70 interactome in cancer.
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.
authors.affiliation: Program in Molecular Pharmacology and Chemistry and Department of Medicine and §Program in Molecular Biology, Proteomics Core, Memorial Sloan-Kettering Cancer Center , New York, New York 10021, United States.
authors.forename: Anna
authors.initials: A
authors.lastname: Rodina
authors.forename: Tony
authors.initials: T
authors.lastname: Taldone
authors.forename: Yanlong
authors.initials: Y
authors.lastname: Kang
authors.forename: Pallav D
authors.initials: PD
authors.lastname: Patel
authors.forename: John
authors.initials: J
authors.lastname: Koren
authors.forename: Pengrong
authors.initials: P
authors.lastname: Yan
authors.forename: Erica M
authors.initials: EM
authors.lastname: DaGama Gomes
authors.forename: Chenghua
authors.initials: C
authors.lastname: Yang
authors.forename: Maulik R
authors.initials: MR
authors.lastname: Patel
authors.forename: Liza
authors.initials: L
authors.lastname: Shrestha
authors.forename: Stefan O
authors.initials: SO
authors.lastname: Ochiana
authors.forename: Cristina
authors.initials: C
authors.lastname: Santarossa
authors.forename: Ronnie
authors.initials: R
authors.lastname: Maharaj
authors.forename: Alexander
authors.initials: A
authors.lastname: Gozman
authors.forename: Marc B
authors.initials: MB
authors.lastname: Cox
authors.forename: Hediye
authors.initials: H
authors.lastname: Erdjument-Bromage
authors.forename: Ronald C
authors.initials: RC
authors.lastname: Hendrickson
authors.forename: Leandro
authors.initials: L
authors.lastname: Cerchietti
authors.forename: Ari
authors.initials: A
authors.lastname: Melnick
authors.forename: Monica L
authors.initials: ML
authors.lastname: Guzman
authors.forename: Gabriela
authors.initials: G
authors.lastname: Chiosis
mesh_terms: D000345:Affinity Labels; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D018840:HSP70 Heat-Shock Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D009369:Neoplasms / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D000345:Affinity Labels; D018840:HSP70 Heat-Shock Proteins