Cholesterol Regulates Assembly of Human Islet Amyloid Polypeptide on Model Membranes

• Published in: Journal of molecular biology 2009-10, Vol.393 (3), p.765-775
• Main Authors: Cho, Won-Jin, Trikha, Saurabh, Jeremic, Aleksandar M.
• Format: Article
• Language: English
• Subjects: AFM; amylin; Amyloid - metabolism; cholesterol; Cholesterol - pharmacology; Humans; Islet Amyloid Polypeptide; membranes; Membranes, Artificial; Microscopy, Atomic Force; Peptides - metabolism; Protein Structure, Quaternary; Solubility - drug effects; Solutions; supramolecular assembly
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2009.08.055

Amylin, a 37-aa pancreatic hormone, is the major constituent of islet amyloid, a hallmark of type II diabetes mellitus. Recent studies have revealed a pivotal role of anionic phospholipids in membrane-catalyzed amylin fibrillogenesis and aggregation. However, cholesterol, an integral component of eukaryotic cell membranes, also could have a role. In this study, we have examined the effect of cholesterol on amylin polymerization both on planar membranes and in solution. Using time-lapse atomic force microscopy, we have studied the dynamics and macromolecular organization of amylin on anionic and neutral planar membranes that lack or include cholesterol. On cholesterol-depleted planar membranes, amylin formed highly symmetrical tetrameric and pentameric pore-like supramolecular structures composed of 25- to 35-nm intermediate-sized globular structures or oligomers. Conversely, on membranes incorporating cholesterol, amylin formed highly compact ∼ 200- to 500-nm protein clusters that constituted seeds or nuclei for continuing amylin binding and aggregation. However, cholesterol inhibited amylin nucleation with a 7-fold decrease in the number of amylin particles. Consequently, cholesterol-containing membranes accumulated significantly less amyloid with some membrane areas completely free of amyloid particles. The inhibitory effect of cholesterol on amylin aggregation in solution was also demonstrated as a 16-fold decrease in the aggregation rate. Consistent with this, circular dichroism spectroscopy revealed a stable, soluble random-coil conformation for amylin in the presence of cholesterol that could explain the inhibitory effect of cholesterol on amylin polymerization in solution and on membranes. The modulatory effect of cholesterol was largely independent of membrane charge or phospholipids, suggesting a novel cholesterol-regulated amylin polymerization process.