dc.contributor.author | Sialana, FJ | |
dc.contributor.author | Roumeliotis, TI | |
dc.contributor.author | Bouguenina, H | |
dc.contributor.author | Chan Wah Hak, L | |
dc.contributor.author | Wang, H | |
dc.contributor.author | Caldwell, J | |
dc.contributor.author | Collins, I | |
dc.contributor.author | Chopra, R | |
dc.contributor.author | Choudhary, JS | |
dc.coverage.spatial | United States | |
dc.date.accessioned | 2022-10-05T13:10:49Z | |
dc.date.available | 2022-10-05T13:10:49Z | |
dc.date.issued | 2022-08-05 | |
dc.identifier.citation | Journal of Proteome Research, 2022, 21 (8), pp. 1842 - 1856 | |
dc.identifier.issn | 1535-3893 | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/5519 | |
dc.identifier.eissn | 1535-3907 | |
dc.identifier.eissn | 1535-3907 | |
dc.identifier.doi | 10.1021/acs.jproteome.2c00092 | |
dc.description.abstract | Large scale proteomic profiling of cell lines can reveal molecular signatures attributed to variable genotypes or induced perturbations, enabling proteogenomic associations and elucidation of pharmacological mechanisms of action. Although isobaric labeling has increased the throughput of proteomic analysis, the commonly used sample preparation workflows often require time-consuming steps and costly consumables, limiting their suitability for large scale studies. Here, we present a simplified and cost-effective one-pot reaction workflow in a 96-well plate format (SimPLIT) that minimizes processing steps and demonstrates improved reproducibility compared to alternative approaches. The workflow is based on a sodium deoxycholate lysis buffer and a single detergent cleanup step after peptide labeling, followed by quick off-line fractionation and MS2 analysis. We showcase the applicability of the workflow in a panel of colorectal cancer cell lines and by performing target discovery for a set of molecular glue degraders in different cell lines, in a 96-sample assay. Using this workflow, we report frequently dysregulated proteins in colorectal cancer cells and uncover cell-dependent protein degradation profiles of seven cereblon E3 ligase modulators (CRL4CRBN). Overall, SimPLIT is a robust method that can be easily implemented in any proteomics laboratory for medium-to-large scale TMT-based studies for deep profiling of cell lines. | |
dc.format | Print-Electronic | |
dc.format.extent | 1842 - 1856 | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | AMER CHEMICAL SOC | |
dc.relation.ispartof | Journal of Proteome Research | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.subject | CELMoDs | |
dc.subject | IMiDs | |
dc.subject | TMTpro | |
dc.subject | cancer cell lines | |
dc.subject | isobaric labeling | |
dc.subject | targeted protein degradation | |
dc.subject | Colorectal Neoplasms | |
dc.subject | Humans | |
dc.subject | Proteome | |
dc.subject | Proteomics | |
dc.subject | Reproducibility of Results | |
dc.subject | Workflow | |
dc.title | SimPLIT: Simplified Sample Preparation for Large-Scale Isobaric Tagging Proteomics. | |
dc.type | Journal Article | |
dcterms.dateAccepted | 2022-07-18 | |
dc.date.updated | 2022-10-05T13:09:37Z | |
rioxxterms.version | VoR | |
rioxxterms.versionofrecord | 10.1021/acs.jproteome.2c00092 | |
rioxxterms.licenseref.startdate | 2022-08-05 | |
rioxxterms.type | Journal Article/Review | |
pubs.author-url | https://www.ncbi.nlm.nih.gov/pubmed/35848491 | |
pubs.issue | 8 | |
pubs.organisational-group | /ICR | |
pubs.organisational-group | /ICR/Primary Group | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Biology | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics/Medicinal Chemistry 2 | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Therapeutics/Medicinal Chemistry 4 (including Analytical Chemistry) | |
pubs.organisational-group | /ICR/Primary Group/ICR Divisions/Cancer Biology/Functional Proteomics Group | |
pubs.organisational-group | /ICR/ImmNet | |
pubs.publication-status | Published | |
pubs.publisher-url | http://dx.doi.org/10.1021/acs.jproteome.2c00092 | |
pubs.volume | 21 | |
icr.researchteam | Functional Proteomics | |
icr.researchteam | Medicinal Chemistry 4 | |
icr.researchteam | Medicinal Chemistry 2 | |
icr.researchteam | Prote & Metabolomics Fac | |
dc.contributor.icrauthor | Sialana, Fernando Jr | |
dc.contributor.icrauthor | Roumeliotis, Theodoros | |
dc.contributor.icrauthor | Wang, Hannah | |
dc.contributor.icrauthor | Caldwell, John | |
dc.contributor.icrauthor | Collins, Ian | |
dc.contributor.icrauthor | Choudhary, Jyoti | |
icr.provenance | Deposited by Mr Arek Surman on 2022-10-05. Deposit type is initial. No. of files: 1. Files: SimPLIT Simplified Sample Preparation for Large-Scale Isobaric Tagging Proteomics.pdf | |