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Mol Neurodegener. 2018 Apr 6;13(1):18. doi: 10.1186/s13024-018-0248-6.

Structural and mechanistic aspects influencing the ADAM10-mediated shedding of the prion protein.

Author information

1
Institute of Neuropathology, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
2
Institute of Biochemistry, Christian Albrechts University, Kiel, Germany.
3
German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany.
4
Division of Molecular Neurobiology, Institute of Enzyme Research, Tokushima University, Tokushima, Japan.
5
Department of Psychiatry and Psychotherapy, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
6
Institute of Biochemistry and Pathobiochemistry, Ruhr University, Bochum, Germany.
7
Institute of Neuropathology, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany. m.glatzel@uke.de.
8
Institute of Neuropathology, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany. h.altmeppen@uke.de.

Abstract

Background:

Proteolytic processing of the prion protein (PrP<superscript>C</superscript>) by endogenous proteases generates bioactive membrane-bound and soluble fragments which may help to explain the pleiotropic roles of this protein in the nervous system and in brain diseases. Shedding of almost full-length PrP<superscript>C</superscript> into the extracellular space by the metalloprotease ADAM10 is of peculiar relevance since soluble PrP stimulates axonal outgrowth and is protective in neurodegenerative conditions such as Alzheimer’s and prion disease. However, molecular determinates and mechanisms regulating the shedding of PrP are entirely unknown.

Methods:

We produced an antibody recognizing the neo-epitope of shed PrP generated by ADAM10 in biological samples and used it to study structural and mechanistic aspects affecting the shedding. For this, we investigated genetically modified cellular and murine models by biochemical and morphological approaches.

Results:

We show that the novel antibody specifically detects shed PrP in cell culture supernatants and murine brain. We demonstrate that ADAM10 is the exclusive sheddase of PrP<superscript>C</superscript> in the nervous system and reveal that the glycosylation state and type of membrane-anchorage of PrP<superscript>C</superscript> severely affect its shedding. Furthermore, we provide evidence that PrP shedding can be modulated by pharmacological inhibition and stimulation and present data suggesting that shedding is a relevant part of a compensatory network ensuring PrP<superscript>C</superscript> homeostasis of the cell.

Conclusions:

With the new antibody, our study introduces a new tool to reliably investigate PrP-shedding. In addition, this study provides novel and important insight into the regulation of this cleavage event, which is likely to be relevant for diagnostic and therapeutic approaches even beyond neurodegeneration.

KEYWORDS:

ADAM10; Antibody; Exosomes; Glycosylation; Membrane anchor; Neurodegeneration; Prion protein; Proteolytic cleavage; Shedding

PMID:
29625583
PMCID:
PMC5889536
DOI:
10.1186/s13024-018-0248-6
[Indexed for MEDLINE]
Free PMC Article

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