4S-fluorination of ProB29 in insulin lispro slows fibril formation

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dc.contributor.authorBreunig, Stephanie L.
dc.contributor.authorChapman, Alex M.
dc.contributor.authorLeBron, Jeanne
dc.contributor.authorQuijano, Janine C.
dc.contributor.authorRanasinghe, Maduni
dc.contributor.authorRawson, Jeffrey
dc.contributor.authorDemeler, Borries
dc.contributor.authorKu, Hsun Teresa
dc.contributor.authorTirrell, David A.
dc.date.accessioned2024-08-19T18:23:04Z
dc.date.available2024-08-19T18:23:04Z
dc.date.issued2024
dc.descriptionOpen access article. Creative Commons Attribution 4.0 International license (CC BY 4.0) applies
dc.description.abstractRecombinant insulin is a life-saving therapeutic for millions of patients affected by diabetes mellitus. Standard mutagenesis has led to insulin variants with improved control of blood glucose; for instance, the fast-acting insulin lispro contains two point mutations that suppress dimer formation and expedite absorption. However, insulins undergo irreversible denaturation, a process accelerated for the insulin monomer. Here we replace ProB29 of insulin lispro with 4R-fluoroproline, 4S-fluoroproline, and 4,4-difluoroproline. All three fluorinated lispro variants reduce blood glucose in diabetic mice, exhibit similar secondary structure as measured by CD, and rapidly dissociate from the zinc- and resorcinol-bound hexamer upon dilution. Notably, however, we find that 4S-fluorination of ProB29 delays the formation of undesired insulin fibrils that can accumulate at the injection site in vivo and can complicate insulin production and storage. These results demonstrate how subtle molecular changes achieved through non-canonical amino acid mutagenesis can improve the stability of protein therapeutics.
dc.description.peer-reviewYes
dc.identifier.citationBreunig, S. L., Chapman, A. M., LeBon, J., Quijano, J. C., Ranasinghe, M., Rawson, J., Demeler, B., Ku, H. T., & Tirrell, D. A. (2024). 4S-fluorination of ProB29 in insulin lispro slows fibril formation. Journal of Biological Chemistry, 399(6), Article. 107332. https://doi.org/10.1016/j.jbc.2024.107332
dc.identifier.urihttps://hdl.handle.net/10133/6865
dc.language.isoen
dc.publisherElsevier
dc.publisher.departmentDepartment of Chemistry and Biochemistry
dc.publisher.facultyArts and Science
dc.publisher.institutionCalifornia Institute of Technology
dc.publisher.institutionBeckman Research Institute City of Hope
dc.publisher.institutionUniversity of Lethbridge
dc.publisher.institutionUniversity of Montana
dc.publisher.institutionIrell & Manella Graduate School of Biological Science
dc.publisher.urlhttps://doi.org/10.1016/j.jbc.2024.107332
dc.subjectNon-canonical amino acid
dc.subjectFluoroproline
dc.subjectProline
dc.subjectInsulin
dc.subjectInsulin lispro
dc.subjectFibrillation
dc.subject4S-fluorination
dc.subjectProtein therapeutics
dc.subject.lcshBlood glucose
dc.subject.lcshInsulin--Therapeutic use
dc.title4S-fluorination of ProB29 in insulin lispro slows fibril formation
dc.typeArticle
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