Insertion of a Synthetic Switch Into Insulin Provides Metabolite-Dependent Regulation of Hormone-Receptor Activation
dc.contributor.author | Chen, Yen-Shan | |
dc.contributor.author | Gleaton, Jeremy | |
dc.contributor.author | Yang, Yanwu | |
dc.contributor.author | Dhayalan, Balamurugan | |
dc.contributor.author | Phillips, Nelson B. | |
dc.contributor.author | Liu, Yule | |
dc.contributor.author | Broadwater, Laurie | |
dc.contributor.author | Jarosinski, Mark | |
dc.contributor.author | Chatterjee, Deepak | |
dc.contributor.author | Lawrence, Michael C. | |
dc.contributor.author | Hattier, Thomas | |
dc.contributor.author | Michael, Dodson M. | |
dc.contributor.author | Weiss, Michael Aaron | |
dc.contributor.department | Biochemistry and Molecular Biology, School of Medicine | en_US |
dc.date.accessioned | 2022-10-25T18:58:53Z | |
dc.date.available | 2022-10-25T18:58:53Z | |
dc.date.issued | 2021-05-03 | |
dc.description.abstract | Insulin signaling requires conformational change: whereas the free hormone and its receptor each adopt autoinhibited conformations, their binding leads to large-scale structural reorganization. To test the coupling between insulin’s “opening” and receptor activation, we inserted an artificial ligand-dependent switch into insulin. Ligand binding disrupts an internal tether designed to stabilize the hormone’s native closed and inactive conformation, thereby enabling productive receptor engagement. This scheme exploited a diol sensor (meta-fluoro-phenylboronic acid at GlyA1) and internal diol (3,4-dihydroxybenzoate at LysB28). The sensor recognizes monosaccharides (fructose > glucose). Studies of insulin signaling in human hepatoma-derived cells (HepG2) demonstrated fructose-dependent receptor autophosphorylation leading to appropriate downstream signaling events, including a specific kinase cascade and metabolic gene regulation (gluconeogenesis and lipogenesis). Addition of glucose (an isomeric ligand with negligible sensor affinity) did not activate the receptor. Similarly, metabolite-regulated signaling was not observed in control studies of (i) an unmodified insulin analog or (ii) an analog containing a diol sensor in the absence of internal tethering. Although as expected CD-detected secondary structure was unaffected by ligand binding, heteronuclear NMR studies revealed subtle local and nonlocal monosaccharide-dependent changes in structure. Insertion of a synthetic switch into insulin has thus demonstrated coupling between hinge-opening and holoreceptor signaling. In addition to this basic finding, our results provide proof of principle for a mechanism-based metabolite-responsive insulin. In particular, replacement of the present fructose sensor by an analogous glucose sensor may enable translational development of a “smart” insulin analog designed to mitigate risk of hypoglycemia in the treatment of diabetes mellitus. | en_US |
dc.eprint.version | Final published version | en_US |
dc.identifier.citation | Chen YS, Gleaton J, Yang Y, et al. Insertion of a Synthetic Switch Into Insulin Provides Metabolite-Dependent Regulation of Hormone-Receptor Activation. J Endocr Soc. 2021;5(Suppl 1):A440. Published 2021 May 3. doi:10.1210/jendso/bvab048.899 | en_US |
dc.identifier.uri | https://hdl.handle.net/1805/30410 | |
dc.language.iso | en_US | en_US |
dc.publisher | Endocrine Society | en_US |
dc.relation.isversionof | 10.1210/jendso/bvab048.899 | en_US |
dc.relation.journal | Journal of the Endocrine Society | en_US |
dc.rights | Attribution-NonCommercial-NoDerivatives 4.0 International | * |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | * |
dc.source | PMC | en_US |
dc.subject | Insulin signaling | en_US |
dc.subject | Ligand binding | en_US |
dc.subject | Monosaccharides | en_US |
dc.subject | Analogous glucose sensor | en_US |
dc.title | Insertion of a Synthetic Switch Into Insulin Provides Metabolite-Dependent Regulation of Hormone-Receptor Activation | en_US |
dc.type | Article | en_US |