{"doi":"10.1101/759662","title":"Generation of Recombinant Mammalian Selenoproteins through Genetic Code Expansion with Photocaged Selenocysteine","abstract":"<jats:title>ABSTRACT</jats:title>\n                <jats:p>\n                  Selenoproteins contain the amino acid selenocysteine and are found in all domains of life. The functions of many selenoproteins are poorly understood, partly due to difficulties in producing recombinant selenoproteins for cell-biological evaluation. Endogenous mammalian selenoproteins are produced through a non-canonical translation mechanism requiring suppression of the UGA stop codon, and a selenocysteine insertion sequence (SECIS) element in the 3’ untranslated region of the mRNA. Here, recombinant selenoproteins are generated in mammalian cells through genetic code expansion, circumventing the requirement for the SECIS element, and selenium availability. An engineered orthogonal\n                  <jats:italic>E. coli</jats:italic>\n                  leucyl-tRNA synthetase/tRNA pair is used to incorporate a photocaged selenocysteine (DMNB-Sec) at the UAG amber stop codon. Recombinantly expressed selenoproteins can be photoactivated in living cells with spatial and temporal control. Using this approach, the native selenoprotein methionine-\n                  <jats:italic>R</jats:italic>\n                  -sulfoxide reductase 1 is generated and activated in mammalian cells. The ability to site-specifically introduce selenocysteine directly in mammalian cells, and temporally modulate selenoprotein activity, will aid in the characterization of mammalian selenoprotein function.\n                </jats:p>","journal":null,"year":null,"id":689066,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"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":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":992119,"name":"Rachel E. Kelemen","orcid":null,"position":1,"is_corresponding":false},{"id":135935,"name":"Masahiro Abo","orcid":null,"position":2,"is_corresponding":false},{"id":1799615,"name":"Laura C. Edinger","orcid":null,"position":3,"is_corresponding":false},{"id":460799,"name":"Jingjia Chen","orcid":"0000-0002-6023-1033","position":4,"is_corresponding":false},{"id":703589,"name":"Abhishek Chatterjee","orcid":"0000-0002-8003-7991","position":5,"is_corresponding":false},{"id":135937,"name":"Eranthie Weerapana","orcid":"0000-0002-0835-8301","position":6,"is_corresponding":false},{"id":1800170,"name":"Jennifer C. Peeler","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Generation of Recombinant Mammalian Selenoproteins through Genetic Code Expansion with Photocaged Selenocysteine","abstract":"<jats:title>ABSTRACT</jats:title>\n                <jats:p>\n                  Selenoproteins contain the amino acid selenocysteine and are found in all domains of life. The functions of many selenoproteins are poorly understood, partly due to difficulties in producing recombinant selenoproteins for cell-biological evaluation. Endogenous mammalian selenoproteins are produced through a non-canonical translation mechanism requiring suppression of the UGA stop codon, and a selenocysteine insertion sequence (SECIS) element in the 3’ untranslated region of the mRNA. Here, recombinant selenoproteins are generated in mammalian cells through genetic code expansion, circumventing the requirement for the SECIS element, and selenium availability. An engineered orthogonal\n                  <jats:italic>E. coli</jats:italic>\n                  leucyl-tRNA synthetase/tRNA pair is used to incorporate a photocaged selenocysteine (DMNB-Sec) at the UAG amber stop codon. Recombinantly expressed selenoproteins can be photoactivated in living cells with spatial and temporal control. Using this approach, the native selenoprotein methionine-\n                  <jats:italic>R</jats:italic>\n                  -sulfoxide reductase 1 is generated and activated in mammalian cells. The ability to site-specifically introduce selenocysteine directly in mammalian cells, and temporally modulate selenoprotein activity, will aid in the characterization of mammalian selenoprotein function.\n                </jats:p>","is_dataset_classified":null,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"26207759","pmcid":null,"openalex_id":"https://openalex.org/W2972005410","authors":[],"funders":[{"funder_name":"National Institutes of Health","grant_id":"5R01GM117004-04","title":"Investigating cysteine PTMs in living cells"},{"funder_name":"National Institutes of Health","grant_id":"1R01GM124319-01","title":"A novel approach to define the roles of oxidative and nitrative post-trasnlational modifications of tryptophan in human biology"},{"funder_name":"National Institutes of Health","grant_id":"5R01GM118431-04","title":"Investigating cysteine-mediated protein activities in C. elegans"},{"funder_name":"National Institutes of Health","grant_id":"1R01GM126220-01A1","title":"A novel strategy to capture post-translational modification-triggered protein-protein interactions"},{"funder_name":"National Institutes of Health","grant_id":"1F32GM131615-01","title":"Effect of FABP5 oxidation on fatty acid binding and EGF signaling"}],"total_grants":5,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[{"year":2019,"count":1}],"oa_status":"green","license":"cc-by-nc","oa_locations":[{"url":"https://www.biorxiv.org/content/biorxiv/early/2019/09/05/759662.full.pdf","host_type":"repository"},{"url":"https://www.biorxiv.org/content/biorxiv/early/2019/09/05/759662.full.pdf","host_type":"repository"},{"url":"https://syndication.highwire.org/content/doi/10.1101/759662","host_type":"publisher"},{"url":"https://doi.org/10.1101/759662","host_type":"repository"},{"url":"https://figshare.com/articles/Generation_of_Recombinant_Mammalian_Selenoproteins_through_Genetic_Code_Expansion_with_Photocaged_Selenocysteine/12257663","host_type":"repository"},{"url":"https://dx.doi.org/10.1101/759662","host_type":""},{"url":"http://dx.doi.org/10.1101/759662","host_type":""}],"fields_of_study":["Selenium in Biological Systems","Organoselenium and organotellurium chemistry","Redox biology and oxidative stress","0301 basic medicine","0303 health sciences","03 medical and health sciences"],"mesh_terms":[],"keywords":["Selenocysteine","Selenoprotein","Stop codon","Genetic code","Transfer RNA","Recombinant DNA","Amino acid","Biology","Biochemistry","Selenoprotein P","Chemistry","Genetics","Gene","Enzyme","RNA","Glutathione","Cysteine","Glutathione peroxidase"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-19T22:01:06.493566Z","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":[]}