Amyloid β-protein (Aβ) has been implicated as an early and essential factor in the pathogenesis of Alzheimer's disease. Although its cellular production has been studied extensively, little is known about Aβ clearance. Recently, insulin-degrading enzyme (IDE), a 110-kDa metalloendopeptidase, was found to degrade both endogenously secreted and synthetic Aβ peptides. Surprisingly, IDE-mediated proteolysis of [125I]Aβ(1-40) in microglial cell-culture media was accompanied by the formation of 125I-labelled peptides with higher apparent molecular masses, raising the possibility that the degradation products act as ‘seeds’ for Aβ oligomerization. To directly address the role of IDE in Aβ degradation and oligomerization, we investigated the action of purified recombinant wild-type and catalytically inactive IDEs. Our data demonstrate that (i) IDE alone is sufficient to cleave purified Aβ that is either unlabelled, iodinated or 35S-labelled; (ii) the initial cleavage sites are His14–Gln15, Phe19–Phe20 and Phe20–Ala21; and (iii) incubation of IDE with [125I]Aβ, but not with [35S]-Aβ, leads to the formation of slower migrating species on gels. Since iodination labels N-terminal fragments of Aβ, and 35S labels C-terminal products, we analysed unlabelled synthetic fragments of Aβ and determined that only the N-terminal fragments migrate with anomalously high molecular mass. These results indicate that IDE alone is sufficient to degrade Aβ at specific sites, and that its degradation products do not promote oligomerization of the intact Aβ peptide.
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October 2000
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Research Article|
October 10 2000
Purified recombinant insulin-degrading enzyme degrades amyloid β-protein but does not promote its oligomerization
Valérie CHESNEAU;
Valérie CHESNEAU
1
*Ben May Institute for Cancer Research, University of Chicago, 5841 S. Maryland Avenue, MC 6027, Chicago, IL 60637, U.S.A.
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Konstantinos VEKRELLIS;
*Ben May Institute for Cancer Research, University of Chicago, 5841 S. Maryland Avenue, MC 6027, Chicago, IL 60637, U.S.A.
2To whom correspondence should be addressed.
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Marsha Rich ROSNER;
Marsha Rich ROSNER
*Ben May Institute for Cancer Research, University of Chicago, 5841 S. Maryland Avenue, MC 6027, Chicago, IL 60637, U.S.A.
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Dennis J. SELKOE
Dennis J. SELKOE
†Center for Neurologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Harvard Institutes of Medicine, 77 Avenue Louis Pasteur, Boston, MA 02115, U.S.A.
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Publisher: Portland Press Ltd
Received:
February 03 2000
Revision Received:
June 14 2000
Accepted:
July 12 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2000
2000
Biochem J (2000) 351 (2): 509–516.
Article history
Received:
February 03 2000
Revision Received:
June 14 2000
Accepted:
July 12 2000
Citation
Valérie CHESNEAU, Konstantinos VEKRELLIS, Marsha Rich ROSNER, Dennis J. SELKOE; Purified recombinant insulin-degrading enzyme degrades amyloid β-protein but does not promote its oligomerization. Biochem J 15 October 2000; 351 (2): 509–516. doi: https://doi.org/10.1042/bj3510509
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