Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 346663
Title Optical microscopy of growing insulin amyloid spherulites on surfaces in vitro
Author(s) Rogers, S.S.; Krebs, M.R.H.; Bromley, E.H.C.; Linden, E. van der; Donald, A.M.
Source Biophysical Journal 90 (2006)3. - ISSN 0006-3495 - p. 1043 - 1054.
DOI https://doi.org/10.1529/biophysj.105.072660
Department(s) Physics and Physical Chemistry of Foods
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2006
Keyword(s) canine mammary-tumors - fibril formation - electron-microscopy - bovine insulin - aggregation - mechanism - evolution - kinetics
Abstract Amyloid fibrils are often found arranged into large ordered spheroid structures, known as spherulites, occurring in vivo and in vitro. The spherulites are predominantly composed of radially ordered amyloid fibrils, which self-assemble from protein in solution. We have observed and measured amyloid spherulites forming from heat-treated solutions of bovine insulin at low pH. The spherulites form in large numbers as semispherical dome-shaped objects on the cell surfaces, showing that surface defects or impurities, or the substrates themselves, can provide good nucleation sites for their formation. Using optical microscopy, we have measured the growth of individual spherulites as a function of time and in various conditions. There is a lag time before nucleation of the spherulites. Once they have nucleated, they grow, each with a radius increasing linearly, or faster than linearly, with time. Remarkably, this growth period has a sudden end, at which all spherulites in the system suddenly stop growing. A model of spherulite formation based on the polymerization of oriented fibrils around a nucleus, from a precursor in solution, quantitatively accounts for the observed growth kinetics. Seeding of native insulin solutions with preformed spherulites led to the preformed spherulites growing without a lag time. This seeding behavior is evidence that the fibrils in the spherulites assemble from small protein species rather than fibrils. The density of the spherulites was also measured and found to be constant with respect to radius, indicating that the space fills as the spherulite grows.
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