Structurally Based Design of Glucagon Mutants That Inhibit Fibril Formation
Name
nihms-1717277.pdf
Description
Accepted version
Size
2.51 MB
Format
Adobe PDF
Checksum (MD5)
7eb6d40542ba7a292f0ff8191e410791
Author(s)
Gelenter, Martin D
Dregni, Aurelio J
Duan, Pu
Hong, Mei
Date Issued
2021
Journal
Biochemistry
Publisher
American Chemical Society (ACS)
Citation
Gelenter, Martin D, Dregni, Aurelio J, Duan, Pu and Hong, Mei. 2021. "Structurally Based Design of Glucagon Mutants That Inhibit Fibril Formation." Biochemistry, 60 (25).
Version
Author's final manuscript
Abstract
The peptide hormone glucagon is prescribed as a pharmaceutical compound to treat diabetic hypoglycemia. However, at the acidic pH where it is highly soluble, glucagon rapidly aggregates into inactive and cytotoxic amyloid fibrils. The recently determined high-resolution structure of these fibrils revealed various stabilizing molecular interactions. On the basis of this structure, we have now designed four arginine mutants of glucagon that resist fibrillization at pharmaceutical concentrations for weeks. An S2R, T29R double mutant and a T29R single mutant remove a hydrogen-bonding interaction in the wild-type fibril, whereas a Y13R, A19R double mutant and a Y13R mutant remove a cation-π interaction. 1H solution nuclear magnetic resonance spectra and ultraviolet absorbance data indicate that these mutants remain soluble in pH 2 buffer under quiescent conditions at concentrations of ≤4 mg/mL for weeks. Under stressed conditions with high salt concentrations and agitation, these mutants fibrillize significantly more slowly than the wild type. The S2R, T29R mutant and the T29R mutant exhibit a mixture of random coil and α-helical conformations, while the Y13R mutant is completely random coil. The mutation sites are chosen to be uninvolved in strong interactions with the glucagon receptor in the active structure of the peptide. Therefore, these arginine mutants of glucagon are promising alternative compounds for treating hypoglycemia.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Creative Commons Attribution-Noncommercial-Share Alike
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1021/ACS.BIOCHEM.1C00214
