Conformational transitions of islet amyloid polypeptide (IAPP) in amyloid formation in vitro

Rakez Kayed, Jürgen Bernhagen, Norma Greenfield, Khuloud Sweimeh, Herwig Brunner, Wolfgang Voelter, Aphrodite Kapurniotu

Research output: Contribution to journalArticlepeer-review

324 Scopus citations


Amyloid aggregates have been recognized to be a pathological hallmark of several fatal diseases, including Alzheimer's disease, the prion-related diseases, and type II diabetes. Pancreatic amyloidosis is characterized by the deposition of amyloid consisting of islet amyloid polypeptide (IAPP). We followed the steps preceding IAPP insolubilization and amyloid formation in vitro using a variety of biochemical methods, including a filtration assay, far and near-UV circular dichroism (CD) spectropolarimetry, 1-anilino-8-naphthalenesulfonic acid (ANS) binding, and atomic force (AFM) and electron (EM) microscopy. IAPP insolubilization and amyloid formation followed kinetics that were consistent with the nucleation-dependent polymerization mechanism. Nucleation of IAPP amyloid formation with traces of preformed fibrils induced a rapid conformational transition into β-sheets that subsequently aggregated into insoluble amyloid fibrils. Transition proceeded via a molten globule-like conformeric state with large contents of secondary structure, fluctuating tertiary and quaternary aromatic interactions, and strongly solvent-exposed hydrophobic patches. In the temperature denaturation pathway at 5 μM peptide, we found that this state was mostly populated at about 45°C, and either aggregated rapidly into amyloid by prolonged exposure to this temperature, or melted into denaturated but still structured IAPP, when heated further to 65°C. The state at 45°C was also found to be populated at 4.25 M GdnHCl at 25°C during GdnHCl-induced equilibrium denaturation, and was stable in solution for several hours before aggregating Into amyloid fibrils. Our studies suggested that this amyloidogenic state was a self-associated form of an aggregation-prone, partially folded state of IAPP. We propose that this partially folded population and its self-associated forms are in a concentration-dependent equilibrium with a non-amyloidogenic IAPP conformer and may act as early, soluble precursors of β-sheet and amyloid formation. Our findings on the molecular mechanism of IAPP amyloid formation in vitro should assist in gaining insight into the pathogenesis and inhibition of pancreatic amyloidosis and other amyloid-related diseases.

Original languageEnglish (US)
Pages (from-to)781-796
Number of pages16
JournalJournal of Molecular Biology
Issue number4
StatePublished - Apr 9 1999
Externally publishedYes


  • Amyloid
  • Circular dichroism
  • Islet amyloid polypeptide (IAPP)
  • Nucleation-dependence
  • Partially folded state

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology


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