{"doi":"10.1101/2024.10.29.620861","title":"Directed evolution of the multicopper oxidase laccase for cell surface proximity labeling and electron microscopy","abstract":"Abstract Enzymes that oxidize aromatic substrates have shown utility in a range of cell-based technologies including live cell proximity labeling (PL) and electron microscopy (EM), but are associated with drawbacks such as the need for toxic H 2 O 2 . Here, we explore laccases as a novel enzyme class for PL and EM in mammalian cells. LaccID, generated via 11 rounds of directed evolution from an ancestral fungal laccase, catalyzes the one-electron oxidation of diverse aromatic substrates using O 2 instead of toxic H 2 O 2 , and exhibits activity selective to the surface plasma membrane of both living and fixed cells. We show that LaccID can be used with mass spectrometry-based proteomics to map the changing surface composition of T cells that engage with tumor cells via antigen-specific T cell receptors. In addition, we use LaccID as a genetically-encodable tag for EM visualization of cell surface features in mammalian cell culture and in the fly brain. Our study paves the way for future cell-based applications of LaccID.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2024,"id":484756,"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":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9425,"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":932929,"name":"Heegwang Roh","orcid":"0000-0002-1549-2437","position":1,"is_corresponding":false},{"id":741977,"name":"David González-Pérez","orcid":"0000-0001-9613-4705","position":2,"is_corresponding":false},{"id":576492,"name":"Mason Mackey","orcid":null,"position":3,"is_corresponding":false},{"id":256492,"name":"Keunyoung Kim","orcid":"0000-0002-4725-3211","position":4,"is_corresponding":false},{"id":669577,"name":"Daniel Hoces","orcid":"0000-0002-1451-5166","position":5,"is_corresponding":false},{"id":242243,"name":"Colleen N. McLaughlin","orcid":"0000-0002-7518-8920","position":6,"is_corresponding":false},{"id":299264,"name":"Stephen Adams","orcid":"0000-0002-4725-2091","position":7,"is_corresponding":false},{"id":6650,"name":"Khanh Hung Nguyen","orcid":"0009-0007-0731-1097","position":8,"is_corresponding":false},{"id":242241,"name":"David J. Luginbuhl","orcid":"0000-0002-2142-780X","position":9,"is_corresponding":false},{"id":242246,"name":"Liqun Luo","orcid":"0000-0001-5467-9264","position":10,"is_corresponding":false},{"id":6645,"name":"Namrata D. Udeshi","orcid":"0000-0001-5312-1402","position":11,"is_corresponding":false},{"id":2226,"name":"Steven A. Carr","orcid":"0000-0002-7203-4299","position":12,"is_corresponding":false},{"id":273968,"name":"Rogelio A. Hernández‐López","orcid":"0000-0003-3968-4908","position":13,"is_corresponding":false},{"id":227589,"name":"Mark H. Ellisman","orcid":"0000-0001-8893-8455","position":14,"is_corresponding":false},{"id":1326931,"name":"Miguel Alcalde","orcid":"0000-0001-6780-7616","position":15,"is_corresponding":false},{"id":242245,"name":"Alice Y. Ting","orcid":"0000-0002-8277-5226","position":16,"is_corresponding":false},{"id":174675,"name":"Song-Yi Lee","orcid":null,"position":0,"is_corresponding":true}],"reference_count":75,"raw_metadata":null,"created_at":"2026-07-19T02:07:42.971417Z","pmid":"39554088","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":[]}