Analysis of Baboon IAPP Provides Insight into Amyloidogenicity and Cytotoxicity of Human IAPP

Abstract The polypeptide hormone islet amyloid polypeptide (IAPP) forms islet amyloid in type 2 diabetes, a process which contributes to pancreatic β-cell dysfunction and death. Not all species form islet amyloid and the ability to do so correlates with the primary sequence. Humans form islet amyloid, but baboon IAPP has not been studied. The baboon peptide differs from human IAPP at three positions containing K1I, H18R, and A25T substitutions. The K1I substitution is a rare example of a replacement in the N-terminal region of amylin. The effect of this mutation on amyloid formation has not been studied, but it reduces the net charge, and amyloid prediction programs suggest that it should increase amyloidogenicity. The A25T replacement involves a non-conservative substitution in a region of IAPP that is believed to be important for aggregation, but the effects of this replacement have not been examined. The H18R point mutant has been previously shown to reduce aggregation in vitro. Baboon amylin forms amyloid on the same time scale as human amylin in vitro and exhibits similar toxicity towards cultured β-cells. The K1I replacement in human amylin slightly reduces toxicity, while the A25T substitution accelerates amyloid formation and enhances toxicity. Photochemical cross-linking reveal that the baboon amylin, like human amylin, forms low order oligomers in the lag phase of amyloid formation. Ion-mobility mass spectrometry reveals broadly similar gas phase collisional cross sections for human and baboon amylin monomers and dimers, with some differences in the arrival time distributions. Pre amyloid oligomers formed by baboon amylin, but not baboon amylin fibers are toxic to cultured β-cells. The toxicity of baboon oligomers and lack of significant detectable toxicity with exogenously added amyloid fibers is consistent with the hypothesis that pre-amyloid oligomers are the most toxic species produced during IAPP amyloid formation.