Preparation of synthetic human islet amyloid polypeptide (IAPP) in a stable conformation to enable study of conversion to amyloid-like fibrils

被引:113
作者
Higham, CE
Jaikaran, ETAS
Fraser, PE
Gross, M
Clark, A
机构
[1] Univ Oxford, Dept Human Anat & Genet, Lab Cellular Endocrinol, Oxford OX1 3QT, England
[2] Radcliffe Infirm, Oxford Ctr Diabet Metab & Endocrinol, Diabet Res Labs, Oxford OX2 6HE, England
[3] Univ Toronto, Dept Med Biophys, Ctr Res Neurodegenerat Dis, Toronto, ON, Canada
[4] Univ Oxford, New Chem Lab, Oxford Ctr Mol Sci, Oxford OX1 3QT, England
基金
英国医学研究理事会; 英国惠康基金;
关键词
islet amyloid polypeptide; fibril; beta-sheet; amyloid;
D O I
10.1016/S0014-5793(00)01287-4
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human synthetic islet amyloid polypeptide (hIAPP) is rapidly converted to beta-sheet conformation and fibrils in aqueous media. Optimal solubility conditions for hIAPP were determined by circular dichroism spectroscopy and transmission electron microscopy, hIAPP in trifluoroethanol or hexafluoro-2-isopropanol (HFIP) diluted in water or phosphate buffer (PB) exhibited random structure which was converted to beta-sheet and fibrils with time, hIAPP, solubilised in HFIP, filtered and lyophilised remained in stable random structure for up to 7 days in water; in PB, insoluble aggregates precipitated from which protofilaments and fibrils formed with time. This suggests that amorphous aggregates of hIAPP could initiate islet amyloidosis in vivo. (C) 2000 Federation of European Biochemical Societies.
引用
收藏
页码:55 / 60
页数:6
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