Document Type
Article - post-print
Publication Date
9-2010
Abstract
The aggregation of the 37-residue polypeptide IAPP, as either insoluble amyloid or as small oligomers, appears to play a direct role in the death of pancreatic β-islet cells in type II diabetes. While IAPP has been known to be the primary component of type II diabetes amyloid, the molecular interactions responsible for this aggregation have not been identified. To identify the aggregation-prone region(s), we constructed a library of randomly generated point mutants of IAPP. This mutant IAPP library was expressed in E. coli as genetic fusions to the reporter protein enhanced green fluorescent protein (EGFP). Because IAPP aggregates rapidly, both independently and when fused to EGFP, the fusion protein does not yield a functional, fluorescent EGFP. However, mutations of IAPP that result in non-amyloidogenic sequences remain soluble and allow EGFP to fold and fluoresce. Using this screen, we identified 22 single mutations, 4 double mutations and 2 triple mutations of IAPP that appear to be less amyloidogenic than wild type human IAPP. A comparison of these sequences suggests residues 13 and 15–17 comprise an additional aggregation-prone region outside of the main amyloidogenic region of IAPP.
Original Publication Citation
Fox A, Snollaerts T, Casanova CE, Calciano A, Nogaj LA, Moffet DA. Selection for Non-Amyloidogenic Mutants of Islet Amyloid Polypeptide (IAPP) Identifies an Extended Region for Amyloidogenicity. Biochemistry. 2010;49(36):7783-7789. doi:10.1021/bi100337p.
Publisher Statement
This is an author-manuscript of an article accepted for publication in Biochemistry. The published version of this record is available online at doi: 10.1021/bi100337p.
Digital Commons @ LMU & LLS Citation
Fox, Ayano; Snollaerts, Thibaut; Casanova, Camille Errecart; Calciano, Anastasia; Nogaj, Luiza A.; and Moffet, David A., "Selection for Non-Amyloidogenic Mutants of Islet Amyloid Polypeptide (IAPP) Identifies an Extended Region for Amyloidogenicity" (2010). Chemistry and Biochemistry Faculty Works. 7.
https://digitalcommons.lmu.edu/chem-biochem_fac/7
