{"doi":"10.1101/2025.10.27.684804","title":"Enhancing enzymatic bioconjugation efficiency via installation of a substrate recruitment domain","abstract":"Enzyme mediated bioconjugation provides a method for easy and rapid formation of protein-protein and protein-small molecule conjugates under mild conditions. Promiscuous enzymes are of particular interest because they can catalyze conjugation reactions on a broad set of substrates. However, this promiscuity carries the risk of undesirable off-target modifications. To mitigate this effect, we used computational design to install a substrate recruitment domain (SRD) onto the promiscuous enzyme, tyrosinase. The redesigned tyrosinase, called D42, preferentially modifies tyrosine residues within the peptide core (core) linked to a 6-amino acid recognition motif/sequence (RS) specific for the SRD. Incorporation of the recognition sequence along with a neighboring tyrosine in peptides or proteins allows for rapid D42-mediated conversion of the tyrosine to an orthoquinone, which can be selectively modified with a variety of nucleophiles. We demonstrate the utility of our design system by rapidly installing cytotoxic molecules on a monoclonal antibody.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":579244,"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":0.9527,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1041578,"name":"Kaylee P. Kuzelka","orcid":"0000-0003-4472-875X","position":1,"is_corresponding":false},{"id":740130,"name":"Jonathan M. Ellis","orcid":"0000-0002-1845-8461","position":2,"is_corresponding":false},{"id":646017,"name":"Zhiyuan Yao","orcid":"0000-0003-4812-9850","position":3,"is_corresponding":false},{"id":877125,"name":"Amelia C. McCue","orcid":"0000-0001-8456-4177","position":4,"is_corresponding":false},{"id":911569,"name":"Rodney Park","orcid":"0000-0003-3824-3916","position":5,"is_corresponding":false},{"id":355232,"name":"Satish K. Nair","orcid":"0000-0003-1790-1334","position":6,"is_corresponding":false},{"id":383841,"name":"Albert A. Bowers","orcid":"0000-0001-8214-7484","position":7,"is_corresponding":false},{"id":114738,"name":"Brian Kuhlman","orcid":"0000-0003-4907-9699","position":8,"is_corresponding":false},{"id":1489469,"name":"Christopher Shelby","orcid":"0009-0000-9294-0881","position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":null,"created_at":"2026-07-19T02:58:30.282164Z","pmid":"41279480","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":[]}