Kad, Neil M, Thomson, N H, Smith, D P, Smith, D A, Radford, S E (2001) Beta(2)-microglobulin and its deamidated variant, N17D form amyloid fibrils with a range of morphologies in vitro. Journal of molecular biology, 313 (3). pp. 559-571. ISSN 0022-2836. (doi:10.1006/jmbi.2001.5071) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:42953)
The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided. | |
Official URL: http://dx.doi.org/10.1006/jmbi.2001.5071 |
Abstract
Amyloid fibrils formed by incubation of recombinant wild-type human beta(2)-microglobulin (beta(2)M) ab initio in vitro at low pH and high ionic strength are short and highly curved. By contrast, fibrils extracted from patients suffering from haemodialysis-related amyloidosis and those formed by seeding growth of the wild-type protein in vitro with fibrils ex vivo are longer and straighter than those previously produced ab initio in vitro. Here we explore the effect of growth conditions on morphology of beta(2)M fibrils formed ab initio in vitro from the wild-type protein, as well as a variant form of beta(2)M in which Asn17 is deamidated to Asp (N17D). We show that deamidation results in significant destabilisation of beta(2)M at neutral pH. Despite this, acidification is still necessary to form amyloid from the mutant protein in vitro. Interestingly, at low pH and low ionic strength long, straight fibrils of recombinant beta(2)M are formed in vitro. The fibrils comprise three distinct morphological types when examined using electron microscopy (EM) and atomic force microscopy (AFM) that vary in periodicity and the number of constituent protofibrils. Using kinetic experiments we suggest that the immature fibrils observed previously do not represent intermediates in the assembly of fully mature amyloid, at least under the conditions studied here.
Item Type: | Article |
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DOI/Identification number: | 10.1006/jmbi.2001.5071 |
Subjects: | Q Science |
Divisions: | Divisions > Division of Natural Sciences > Biosciences |
Depositing User: | Neil Kad |
Date Deposited: | 15 Sep 2014 19:07 UTC |
Last Modified: | 05 Nov 2024 10:27 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/42953 (The current URI for this page, for reference purposes) |
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