{"doi":"10.1101/2025.01.06.631444","title":"Optogenetic control of Protein Kinase C-epsilon activity reveals its intrinsic signaling properties with spatiotemporal resolution","abstract":"The regulation of PKC epsilon (PKCε) and its downstream effects is still not fully understood, making it challenging to develop targeted therapies or interventions. A more precise tool that enables spatiotemporal control of PKCε activity is thus required. Here, we describe a photo-activatable optogenetic PKCε probe (Opto-PKCε) consisting of an engineered PKCε catalytic domain and a blue-light inducible dimerization domain. Molecular dynamics and AlphaFold simulations enable rationalization of the dark-light activity of the optogenetic probe. We first characterize the binding partners of Opto-PKCε, which are similar to those of PKCε. Subsequent validation of the Opto-PKCε tool is performed with phosphoproteome analysis, which reveals that only PKCε substrates are phosphorylated upon light activation. Opto-PKCε could be engineered for recruitment to specific subcellular locations. Activation of Opto-PKCε in isolated hepatocytes reveals its sustained activation at the plasma membrane is required for its phosphorylation of the insulin receptor at Thr1160. In addition, Opto-PKCε recruitment to the mitochondria results in its lowering of the spare respiratory capacity through phosphorylation of complex I NDUFS4. These results demonstrate that Opto-PKCε may have broad applications for the studies of PKCε signaling with high specificity and spatiotemporal resolution.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":557012,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9608,"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":1262765,"name":"Crystal Jing Yi Lim","orcid":null,"position":1,"is_corresponding":false},{"id":1262339,"name":"Yilie Liao","orcid":"0009-0008-7122-8690","position":2,"is_corresponding":false},{"id":1189679,"name":"Justin Ng","orcid":"0000-0002-4730-1877","position":3,"is_corresponding":false},{"id":1262338,"name":"Ler Ting Rachel Lim","orcid":"0009-0002-6681-7614","position":4,"is_corresponding":false},{"id":1457161,"name":"S S Koh","orcid":null,"position":5,"is_corresponding":false},{"id":1262764,"name":"Sher En Chan","orcid":null,"position":6,"is_corresponding":false},{"id":1457162,"name":"P Lee","orcid":null,"position":7,"is_corresponding":false},{"id":1457163,"name":"Hui-Jun Lim","orcid":null,"position":8,"is_corresponding":false},{"id":700855,"name":"Chen Ye","orcid":"0000-0003-3935-1601","position":9,"is_corresponding":false},{"id":873100,"name":"Loo Chien Wang","orcid":null,"position":10,"is_corresponding":false},{"id":1262768,"name":"Siok Ghee Ler","orcid":null,"position":11,"is_corresponding":false},{"id":872267,"name":"Radoslaw M. Sobota","orcid":"0000-0002-2455-2526","position":12,"is_corresponding":false},{"id":1244299,"name":"Yaw Sing Tan","orcid":"0000-0002-2522-9421","position":13,"is_corresponding":false},{"id":232241,"name":"Gerald I. Shulman","orcid":"0000-0003-1529-5668","position":14,"is_corresponding":false},{"id":307401,"name":"Xiaoyong Yang","orcid":"0000-0001-7263-3913","position":15,"is_corresponding":false},{"id":572425,"name":"Weiping Han","orcid":"0000-0002-5023-2104","position":16,"is_corresponding":false},{"id":281125,"name":"Qunxiang Ong","orcid":"0000-0003-3674-2105","position":0,"is_corresponding":true}],"reference_count":1,"raw_metadata":null,"created_at":"2026-07-19T02:55:13.130091Z","pmid":"39829776","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":[]}