{"doi":"10.1101/2022.06.01.494336","title":"PPIX-binding Proteins Reveal Porphyrin Synthesis and Ferroptosis Link","abstract":"Abstract All aerobic organisms require the cofactor heme to survive, but its synthesis requires formation of a potentially toxic intermediate protoporphyrin IX (PPIX). Little is known about the extent of PPIX’s cellular interactions. Here, we report the development of PPB, a biotin-conjugated, PPIX-probe that captures proteins capable of interacting with PPIX. Quantitative proteomics with PPB identified common proteins among a diverse panel of mammalian cell lineages. Pathway and quantitative difference analysis revealed PPB-bound proteins related to iron and heme metabolism and suggested that these processes might be altered by heme/porphyrin synthesis. We show that increased heme/porphyrin synthesis in cells promotes ferroptosis that is pharmacologically distinct from canonical ferroptosis driven by erastin, an inhibitor of the cystine/glutamate antiporter. Proteomic data derived from PPB revealed an interactor, PRDX3, a mitochondrial peroxidase, that modulated heme/porphyrin biosynthesis driven ferroptosis. Consistent with a role in porphyrin-induced ferroptotic death targeted gene knockdown of PRDX3, but not peroxidases, PRDX1 or 2, enhanced porphyrin-induced ferroptotic death. The relationship between increased heme/porphyrin synthesis and ferroptosis was also found in a ferrochelatase-deficient T-lymphoblastoid leukemia cell line, suggesting potential strategy for treating certain cancers. We demonstrate that when the PPB probe is coupled with unbiased proteomics a previously unreported relationship between heme/porphyrin synthesis, and ferroptosis was discovered.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":309196,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9551,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1004257,"name":"Yao Wang","orcid":"0000-0001-6672-9601","position":1,"is_corresponding":false},{"id":94638,"name":"Yuxin Li","orcid":"0000-0001-9739-9860","position":2,"is_corresponding":false},{"id":714596,"name":"Kanisha Kavdia","orcid":null,"position":3,"is_corresponding":false},{"id":721835,"name":"Yu Fukuda","orcid":"0000-0001-6182-540X","position":4,"is_corresponding":false},{"id":503693,"name":"Sabina Ranjit","orcid":null,"position":5,"is_corresponding":false},{"id":267294,"name":"Camenzind G. Robinson","orcid":"0000-0002-7277-692X","position":6,"is_corresponding":false},{"id":556039,"name":"Christy R. Grace","orcid":"0000-0002-1600-5610","position":7,"is_corresponding":false},{"id":1004258,"name":"Youlin Xia","orcid":"0000-0001-7255-5425","position":8,"is_corresponding":false},{"id":97162,"name":"Junmin Peng","orcid":"0000-0003-0472-7648","position":9,"is_corresponding":false},{"id":535479,"name":"John D. Schuetz","orcid":"0000-0003-0779-1303","position":10,"is_corresponding":false},{"id":401965,"name":"J.A. Lynch","orcid":"0000-0003-3624-2741","position":0,"is_corresponding":true}],"reference_count":88,"raw_metadata":null,"created_at":"2026-07-19T00:33:11.579763Z","pmid":null,"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":[]}