{"doi":"10.1021/acschembio.3c00570","title":"A Tandem-Affinity Purification Method for Identification of Primary Intracellular Drug-Binding Proteins","abstract":"In the field of drug discovery, understanding how small molecule drugs interact with cellular components is crucial. Our study introduces a novel methodology to uncover primary drug targets using T andem A ffinity P urification for identification of D rug- B inding P roteins (TAP-DBP). Central to our approach is the generation of a FLAG-hemagglutinin (HA)-tagged chimeric protein featuring the FKBP12(F36V) adaptor protein and the TurboID enzyme. Conjugation of drug molecules with the FKBP12(F36V) ligand allows for the coordinated recruitment of drug-binding partners effectively enabling in-cell TurboID-mediated biotinylation. By employing a tandem affinity purification protocol based on FLAG-immunoprecipitation and streptavidin pulldown, alongside mass spectrometry analysis, TAP-DBP allows for the precise identification of drug-primary binding partners. Overall, this study introduces a systematic, unbiased method for identification of drug–protein interactions, contributing a clear understanding of target engagement and drug selectivity to advance the mode of action of a drug in cells.","journal":"ACS Chemical Biology","year":2024,"id":459146,"datarank":0.24141568686511508,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.0,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9618,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1019653,"name":"Jitendra Gour","orcid":"0000-0003-0125-2253","position":1,"is_corresponding":false},{"id":293665,"name":"Thomas Beer","orcid":null,"position":2,"is_corresponding":false},{"id":258003,"name":"Hsin‐Yao Tang","orcid":"0000-0003-1838-018X","position":3,"is_corresponding":false},{"id":348572,"name":"Joel Cassel","orcid":"0000-0001-8465-8739","position":4,"is_corresponding":false},{"id":348575,"name":"Joseph M. Salvino","orcid":"0000-0002-2184-5980","position":5,"is_corresponding":false},{"id":276935,"name":"Luca Busino","orcid":"0000-0001-6758-9276","position":6,"is_corresponding":false},{"id":1071667,"name":"Sehbanul Islam","orcid":"0000-0003-2686-2787","position":0,"is_corresponding":true}],"reference_count":43,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:03:55.280882Z","pmid":"38271588","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}