{"doi":"10.3233/jpd-213128","title":"Evaluation of Current Methods to Detect Cellular Leucine-Rich Repeat Kinase 2 (LRRK2) Kinase Activity","abstract":"BACKGROUND: Coding variation in the Leucine rich repeat kinase 2 gene linked to Parkinson's disease (PD) promotes enhanced activity of the encoded LRRK2 kinase, particularly with respect to autophosphorylation at S1292 and/or phosphorylation of the heterologous substrate RAB10. OBJECTIVE: To determine the inter-laboratory reliability of measurements of cellular LRRK2 kinase activity in the context of wildtype or mutant LRRK2 expression using published protocols. METHODS: Benchmark western blot assessments of phospho-LRRK2 and phospho-RAB10 were performed in parallel with in situ immunological approaches in HEK293T, mouse embryonic fibroblasts, and lymphoblastoid cell lines. Rat brain tissue, with or without adenovirus-mediated LRRK2 expression, and human brain tissues from subjects with or without PD, were also evaluated for LRRK2 kinase activity markers. RESULTS: Western blots were able to detect extracted LRRK2 activity in cells and tissue with pS1292-LRRK2 or pT73-RAB10 antibodies. However, while LRRK2 kinase signal could be detected at the cellular level with over-expressed mutant LRRK2 in cell lines, we were unable to demonstrate specific detection of endogenous cellular LRRK2 activity in cell culture models or tissues that we evaluated. CONCLUSION: Further development of reliable methods that can be deployed in multiple laboratories to measure endogenous LRRK2 activities are likely required, especially at cellular resolution.","journal":"Journal of Parkinson s Disease","year":2022,"id":269036,"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":17,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9584,"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":362347,"name":"Vinita G. Chittoor-Vinod","orcid":"0000-0003-0518-8770","position":1,"is_corresponding":false},{"id":236477,"name":"Jillian H. Kluss","orcid":"0000-0002-6262-6661","position":2,"is_corresponding":false},{"id":307807,"name":"Kaela Kelly","orcid":"0000-0002-3313-9768","position":3,"is_corresponding":false},{"id":688234,"name":"Nicole Bryant","orcid":"0009-0004-7827-9205","position":4,"is_corresponding":false},{"id":309799,"name":"An Phu Tran Nguyen","orcid":null,"position":5,"is_corresponding":false},{"id":304230,"name":"Syed Bukhari","orcid":"0000-0002-6861-2623","position":6,"is_corresponding":false},{"id":297633,"name":"Nathan Smith","orcid":"0000-0002-9837-9957","position":7,"is_corresponding":false},{"id":931753,"name":"Antonio Jesús Lara Ordóñez","orcid":"0000-0002-8884-4886","position":8,"is_corresponding":false},{"id":931754,"name":"Elena Fdez","orcid":"0000-0002-9971-848X","position":9,"is_corresponding":false},{"id":34703,"name":"Marie‐Christine Chartier‐Harlin","orcid":"0000-0001-6416-6526","position":10,"is_corresponding":false},{"id":107086,"name":"Thomas J. Montine","orcid":"0000-0002-1346-2728","position":11,"is_corresponding":false},{"id":388663,"name":"Mark A. Wilson","orcid":"0000-0001-6317-900X","position":12,"is_corresponding":false},{"id":297155,"name":"Darren J. Moore","orcid":"0000-0003-2359-5679","position":13,"is_corresponding":false},{"id":307808,"name":"Andrew B. West","orcid":"0000-0002-3034-4061","position":14,"is_corresponding":false},{"id":238914,"name":"Mark Cookson","orcid":null,"position":15,"is_corresponding":false},{"id":571597,"name":"R. Jeremy Nichols","orcid":"0000-0002-5228-7571","position":16,"is_corresponding":false},{"id":931755,"name":"Sabine Hilfiker","orcid":"0000-0002-5167-7682","position":17,"is_corresponding":false},{"id":931752,"name":"Belén Fernández","orcid":"0000-0001-9987-1495","position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":null,"created_at":"2026-07-19T00:27:18.142851Z","pmid":"35599495","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":[]}