Native Size-Exclusion Chromatography-Based Mass Spectrometry Reveals New Components of the Early Heat Shock Protein 90 Inhibition Response Among Limited Global Changes.

Autor:	Samant RS; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom; Signalling Programme, The Babraham Institute, Cambridge, United Kingdom. Electronic address: rahul.samant@babraham.ac.uk., Batista S; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom., Larance M; Centre for Gene Regulation & Expression, University of Dundee, Dundee, United Kingdom., Ozer B; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom., Milton CI; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom., Bludau I; Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany., Wu E; Signalling Programme, The Babraham Institute, Cambridge, United Kingdom., Biggins L; Bioinformatics Group, The Babraham Institute, Cambridge, United Kingdom., Andrews S; Bioinformatics Group, The Babraham Institute, Cambridge, United Kingdom., Hervieu A; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom., Johnston HE; Signalling Programme, The Babraham Institute, Cambridge, United Kingdom., Al-Lazikhani B; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom; Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA., Lamond AI; Centre for Gene Regulation & Expression, University of Dundee, Dundee, United Kingdom., Clarke PA; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom., Workman P; Centre for Cancer Drug Discovery, The Institute of Cancer Research, London, United Kingdom. Electronic address: paul.workman@icr.ac.uk.
Jazyk:	angličtina
Zdroj:	Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2023 Feb; Vol. 22 (2), pp. 100485. Date of Electronic Publication: 2022 Dec 20.
DOI:	10.1016/j.mcpro.2022.100485
Abstrakt:	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 early changes in native protein complexes following treatment with the HSP90 inhibitor tanespimycin (17-AAG) for 8 h 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 HT29 cells at this relatively early time-point. 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 known and unknown components of the HSP90-dependent proteome. We validate two of these-the actin-binding protein Anillin and the mitochondrial isocitrate dehydrogenase 3 complex-as novel HSP90 inhibitor-modulated proteins. 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. Competing Interests: Conflict of interest The Institute of Cancer Research has a commercial interest in HSP90 and HSF1 inhibitors and operates a reward to discoverers scheme from which employees may benefit. P. W. received funding from Vernalis for the discovery of HSP90 inhibitors, and intellectual property for this program was licensed to Vernalis Ltd and Novartis. P. W. 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. P. W. has been/is a consultant/advisory board member to Alterome Therapeutics, Astex Therapeutics, Black Diamond Therapeutics, CHARM Therapeutics, CV6 Therapeutics, EpiCombi Therapeutics, Novartis, STORM Therapeutics, and Vividion Therapeutics (acquired by Bayer AG) and is a Science Partner for Nextech Invest. P. W. is a Non-Executive Board member and holds stock in STORM Therapeutics and also holds stock in Alterome Therapeutics, Black Diamond Therapeutics, Chroma Therapeutics, EpiCombi Therapeutics, Nextech Invest, and Nuvectis Pharma. P. W. is the Executive Director of the non-profit Chemical Probes Portal. P. W. and P. A. C. received research funding from Merck KGaA and Astex Therapeutics, and P. W. received research funding from AstraZeneca, Battle Against Cancer Investment Trust (BACIT), and CRT Pioneer Fund/Sixth Element Capital). P. W. is a former employee of AstraZeneca. B. A. L. 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). B. A. L. has financial interest and/or acts/acted as a consultant or a Scientific Advisory Board member for Exscientia AI, AstraZeneca, Astex Pharmaceuticals, GSK, Astellas Pharma, and Definiens AG (member of AstraZeneca group). B. A. L. is the Director of Informatics for the non-profit Chemical Probes Portal. (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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