Abstract
The molecular chaperone heat shock protein 90 (HSP90) works in concert with co-chaperones to stabilize its client proteins, which include multiple drivers of oncogenesis and malignant progression. Pharmacologic inhibitors of HSP90 have been observed to exert a wide range of effects on the proteome, including depletion of client proteins, induction of heat shock proteins, dissociation of co-chaperones from HSP90, disruption of client protein signaling networks, and recruitment of the protein ubiquitylation and degradation machinery—suggesting widespread remodeling of cellular protein complexes. However, proteomics studies to date have focused on inhibitor-induced changes in total protein levels, often overlooking protein complex alterations. Here, we use size-exclusion chromatography in combination with mass spectrometry (SEC-MS) to characterize the changes in native protein complexes following treatment with the HSP90 inhibitor tanespimycin (17-AAG) in the HT29 colon adenocarcinoma cell line. After confirming the signature cellular response to HSP90 inhibition (e.g., induction of heat shock proteins, decreased total levels of client proteins), we were surprised to find only modest perturbations to the global distribution of protein elution profiles in inhibitor-treated cells. Similarly, co-chaperones that co-eluted with HSP90 displayed no clear difference between control and treated conditions. However, two distinct analysis strategies identified multiple inhibitor-induced changes, including several known components of the HSP90 proteome, as well as numerous proteins and protein complexes with no previous links to HSP90. We present this dataset as a resource for the HSP90, proteostasis, and cancer communities (https://www.bioinformatics.babraham.ac.uk/shiny/HSP90/SEC-MS/), laying the groundwork for future mechanistic and therapeutic studies related to HSP90 pharmacology. Data are available via ProteomeXchange with identifier PXD033459.
- molecular chaperone
- heat shock protein
- protein complexes
- proteomics
- HSP90 inhibitor
- tanespimycin
Competing Interest Statement
The Institute of Cancer Research has a commercial interest in HSP90 and HSF1 inhibitors and operates a reward to inventors scheme form which employees may benefit. PW received funding from Vernalis for the discovery of HSP90 inhibitors, and intellectual property for this program was licensed to Vernalis Ltd and Novartis. PW was previously involved in a research collaboration with AstraZeneca in the area of the HSF1 pathway and intellectual property was licensed to Sixth Element Capital/Pioneer Fund and Nuvectis Pharma. PW has been/is a consultant/ advisory board member to Alterome Therapeutics, Astex Therapeutics, Black Diamond Therapeutics, CV6 Therapeutics, Novartis, Storm Therapeutics, Vividion Therapeutics (acquired by Bayer AG), and is a Science Partner for Nextechinvest. PW is Non-Executive Board member and holds stock in Storm Therapeutics, and also holds stock in Alterome Therapeutics, Chroma Therapeutics, EpiCombi Therapeutics, NextTechInvest, and Nuvectis Pharma. PW is Executive Director of the non-profit Chemical Probes Portal. PW and PAC received research funding from Merck KGaA and Astex Therapeutics and PW received research funding from AstraZeneca, Battle Against Cancer Investment Trust (BACIT), and CRT Pioneer Fund/Sixth Element Capital). BAL is a former employee of The Institute of Cancer Research which operates reward to inventors program and a former employee of Inpharmatica Ltd (later acquired by Galapagos). PW is a former employee of AstraZeneca. BAL has financial interest and/or acts/acted as consultant or Scientific Advisory Board member for Exscientia AI; AstraZeneca; Astex Pharmaceuticals; GSK; Astellas Pharma; Definiens AG (member of AstraZeneca group). BAL is Director of Informatics for the non-profit Chemical Probes Portal.
