{"doi":"10.1101/2023.06.24.546337","title":"Se-Glargine II. Native Function of a Basal Insulin Analog Stabilized by an Internal Diselenide Bridge","abstract":"Insulin glargine, the active component of basal clinical pharmaceutical formulations Lantus® and Toujeo® (Sanofi), provides a model for principles of therapeutic protein design. Formulated in solution at pH 4, insulin glargine exhibits a shifted isoelectric point (from pH 5.3 to neutral pH) due to a basic dipeptide B-chain extension (Arg B31 -Arg B32 ). In the first article in this series, we described pairwise substitution of Cys A6 and Cys A11 by seleno-cysteine (Sec; the 21 st encoded amino acid) by solid-phase peptide synthesis. 1 H- 2 H amide proton exchange, as monitored by 1 H-NMR spectroscopy, provides evidence that substitution of internal cystine A6-A11 by a diselenide bridge stabilizes the protein and damps segmental conformational fluctuations. Further, this analog and its major metabolites M1 and M2 (respectively denoting proteolytic derivatives lacking Arg B31 -Arg B32 or Thr B30 -Arg B31 -Arg B32 ) exhibit native hormonal activity in mammalian cell-based assays measuring dose-dependent autophosphorylation of the insulin receptor (pIR/IR ratio) and metabolic gene regulation in human liver-derived HepG2 cells. The internal diselenide bridge also did not alter respective baseline mitogenicities of insulin glargine or its proteolytic products as evaluated by a qPCR-based assay of the balance between proliferative and antiproliferative cyclin gene expression; the assays employed L6 myoblasts over-expressing mitogenic IR isoform A. Given such native function, shelf life—and hence global access to insulin in the developing world—may be enhanced by stabilizing diselenide chemistry.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":393405,"datarank":0.24141568686511508,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.0,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":1,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9537,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":338985,"name":"Balamurugan Dhayalan","orcid":"0000-0001-6002-1207","position":1,"is_corresponding":false},{"id":338984,"name":"Yanwu Yang","orcid":"0000-0002-4832-2987","position":2,"is_corresponding":false},{"id":279958,"name":"Orit Weil‐Ktorza","orcid":null,"position":3,"is_corresponding":false},{"id":277901,"name":"Norman Metanis","orcid":"0000-0002-6373-9318","position":4,"is_corresponding":false},{"id":338990,"name":"Michael A. Weiss","orcid":"0000-0003-1325-1610","position":5,"is_corresponding":false},{"id":338987,"name":"Yen‐Shan Chen","orcid":"0000-0003-4746-6636","position":0,"is_corresponding":true}],"reference_count":40,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:19:05.913428Z","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":[]}