Suppression of IAPP fibrillation at anionic lipid membranes via IAPP-derived amyloid inhibitors and insulin.
Aggregation of human islet amyloid polypeptide (hIAPP) into cytotoxic beta-sheet oligomers and amyloid plaques is considered a key event in pancreatic beta-cell degeneration in type 2 diabetes (T2D). hIAPP is synthesized in the pancreatic beta-cells and it is stored, co-processed in the secretory granules, and co-secreted to the extracellular matrix together with insulin. In vivo, hIAPP aggregation may start and proceed at the water-cell membrane interface and anionic lipid membranes strongly enhance the process of hIAPP fibrillization which is causally linked to membrane disintegration and cell degeneration. In this study we explored the amyloidogenic propensity and conformational properties of hIAPP in the presence of negatively charged membrane (DOPC/DOPG phospholipid bilayers) surfaces upon addition of two recently designed potent hIAPP-derived inhibitors of hIAPP amyloidogenesis, the hexapeptide NF(N-Me)GA(N-Me)IL (NFGAIL-GI) and the 37-residue non-amyloidogenic hIAPP analog [(N-Me)G24, (N-Me)I26]-IAPP (IAPP-GI). For comparison, the effects of insulin, which is a natively occurring hIAPP aggregation inhibitor, rat IAPP (rIAPP), which is a natively non-amyloidogenic hIAPP analog, and the hIAPP amyloid core peptide hIAPP(22-27) or NFGAIL were also studied. The aim of our study was to test whether and how the above peptides which have been shown to completely block or suppress hIAPP amyloidogenesis in bulk solution in vitro would also affect these processes in the presence of lipid membranes. To this end, attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) was applied. We find that IAPP-GI, NFGAIL-GI, insulin, and rIAPP are potent inhibitors of hIAPP fibrillization. Importantly, our data also suggest that the hetero-complexes of IAPP-GI, rIAPP, and insulin with hIAPP although non-amyloidogenic per se are still able to adsorb at the lipid membrane. By contrast, in the presence of NFGAIL-GI, interaction of hIAPP with the lipid membrane is completely abolished, consistent with NFGAIL-GI mediated sequestration of hIAPP via hetero-complexation in the aqueous phase mainly accounting for the observed strong effect of NFGAIL-GI on hIAPP fibrillogenesis at the lipid membrane interface. Finally, our studies show that once hIAPP is fibrillized at the water-lipid membrane interface with fibrils being attached to the lipid membrane, it cannot be disaggregated by all above peptides.
Physical Chemistry I - Biophysical Chemistry, Department of Chemistry, TU Dortmund University, Otto-Hahn Str. 6, D-44227, Dortmund, Germany.
Daniel
D
Sellin
Li-Mei
LM
Yan
Aphrodite
A
Kapurniotu
Roland
R
Winter
D000595:Amino Acid Sequence; D000682:Amyloid / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D017550:Spectroscopy, Fourier Transform Infrared
C509234:(N-Me)G24, (N-Me)I26-IAPP; D000682:Amyloid; D007328:Insulin; D008051:Lipid Bilayers; D010455:Peptides; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol; C017251:1,2-oleoylphosphatidylcholine