Mutational analysis of the ability of resveratrol to inhibit amyloid formation by islet amyloid polypeptide

Critical evaluation of the importance of aromatic-inhibitor and histidine-inhibitor interactions

Ling-Hsien Tu, Lydia M. Young, Amy G. Wong, Alison E. Ashcroft, Sheena E. Radford, Daniel P. Raleigh

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The process of amyloid formation by the normally soluble hormone islet amyloid polypeptide (IAPP) contributes to β-cell death in type 2 diabetes and in islet transplants. There are no clinically approved inhibitors of islet amyloidosis, and the mode of action of existing inhibitors is not well-understood. Resveratrol, a natural polyphenol, has been reported to inhibit amyloid formation by IAPP and by the Alzheimer's disease Aβ peptide. The mechanism of action of this compound is not known, nor is its mode of interaction with IAPP. In this study, we use a series of IAPP variants to examine possible interactions between resveratrol and IAPP. Fluorescence assays, transmission electron microscopy, and mass spectrometry demonstrate that resveratrol is much less effective as an inhibitor of IAPP amyloid formation than the polyphenol (-)-epigallocatechin 3-gallate (EGCG) and, unlike EGCG, does not significantly disaggregate preformed IAPP amyloid fibrils. Resveratrol is also shown to interfere with thioflavin-T assays. His-18 mutants, a truncation mutant, mutants of each of the aromatic residues, and mutants of Arg-11 of IAPP were examined. Mutation of His to Gln or Leu weakens the ability of resveratrol to inhibit amyloid formation by IAPP, as do mutations of Arg-11, Phe-15, or Tyr-37 to Leu, and truncation to form the variant Ac 8-37-IAPP, which removes the first seven residues to eliminate Lys-1 and the N-terminal amino group. In contrast, replacement of Phe-23 with Leu has a smaller effect. The data highlight Phe-15, His-18, and Tyr-37 as being important for IAPP-resveratrol interactions and are consistent with a potential role of the N-terminus and Arg-11 in polypeptide-resveratrol interactions. (Graph Presented).

Original languageEnglish
Pages (from-to)666-676
Number of pages11
JournalBiochemistry
Volume54
Issue number3
DOIs
Publication statusPublished - 2015 Jan 27

Fingerprint

Islet Amyloid Polypeptide
Aptitude
Histidine
Amyloid
Polyphenols
Assays
resveratrol
Transplants
Peptides
Mutation
Amyloidosis
Cell death
Medical problems
Transmission Electron Microscopy
Type 2 Diabetes Mellitus
Mass spectrometry
Mass Spectrometry
Alzheimer Disease
Cell Death
Fluorescence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mutational analysis of the ability of resveratrol to inhibit amyloid formation by islet amyloid polypeptide : Critical evaluation of the importance of aromatic-inhibitor and histidine-inhibitor interactions. / Tu, Ling-Hsien; Young, Lydia M.; Wong, Amy G.; Ashcroft, Alison E.; Radford, Sheena E.; Raleigh, Daniel P.

In: Biochemistry, Vol. 54, No. 3, 27.01.2015, p. 666-676.

Research output: Contribution to journalArticle

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abstract = "The process of amyloid formation by the normally soluble hormone islet amyloid polypeptide (IAPP) contributes to β-cell death in type 2 diabetes and in islet transplants. There are no clinically approved inhibitors of islet amyloidosis, and the mode of action of existing inhibitors is not well-understood. Resveratrol, a natural polyphenol, has been reported to inhibit amyloid formation by IAPP and by the Alzheimer's disease Aβ peptide. The mechanism of action of this compound is not known, nor is its mode of interaction with IAPP. In this study, we use a series of IAPP variants to examine possible interactions between resveratrol and IAPP. Fluorescence assays, transmission electron microscopy, and mass spectrometry demonstrate that resveratrol is much less effective as an inhibitor of IAPP amyloid formation than the polyphenol (-)-epigallocatechin 3-gallate (EGCG) and, unlike EGCG, does not significantly disaggregate preformed IAPP amyloid fibrils. Resveratrol is also shown to interfere with thioflavin-T assays. His-18 mutants, a truncation mutant, mutants of each of the aromatic residues, and mutants of Arg-11 of IAPP were examined. Mutation of His to Gln or Leu weakens the ability of resveratrol to inhibit amyloid formation by IAPP, as do mutations of Arg-11, Phe-15, or Tyr-37 to Leu, and truncation to form the variant Ac 8-37-IAPP, which removes the first seven residues to eliminate Lys-1 and the N-terminal amino group. In contrast, replacement of Phe-23 with Leu has a smaller effect. The data highlight Phe-15, His-18, and Tyr-37 as being important for IAPP-resveratrol interactions and are consistent with a potential role of the N-terminus and Arg-11 in polypeptide-resveratrol interactions. (Graph Presented).",
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