Benseny-Cases, N, Karamanos, TK, Hoop, CL et al. (2 more authors) (2019) Extracellular matrix components modulate different stages in β2-microglobulin amyloid formation. The Journal of biological chemistry, 294. pp. 9392-9401. ISSN 0021-9258
Abstract
Amyloid deposition of wild-type human β2-microglobulin (WT-hβ2m) in the joints of long-term hemodialysis patients is the hallmark of dialysis-related amyloidosis (DRA). In vitro, WT-hβ2m does not form amyloid fibrils at physiological pH and temperature unless co-solvents or other reagents are added. Therefore, understanding how fibril formation is initiated and maintained in the joint space is important for elucidating WT-hβ2m aggregation and DRA onset. Here, we investigated the roles of collagen I and the commonly administered anticoagulant, low-molecular-weight (LMW) heparin, in the initiation and subsequent aggregation phases of WT-hβ2m in physiologically relevant conditions. Using thioflavin T (ThT) fluorescence to study the kinetics of amyloid formation, we analyzed how these two agents affect specific stages of WT-hβ2m assembly. Our results revealed that LMW-heparin strongly promotes WT-hβ2m fibrillogenesis during all stages of aggregation. However, collagen I affected WT-hβ2m amyloid formation in contrasting ways: decreasing the lag time of fibril formation in the presence of LMW-heparin and slowing the rate at higher concentrations. We found that in self-seeded reactions, interaction of collagen I with WT-hβ2m amyloid fibrils attenuates surface-mediated growth of WT-hβ2m fibrils, demonstrating a key role of secondary nucleation in WT-hβ2m amyloid formation. Interestingly, collagen I fibrils did not suppress surface-mediated assembly of WT-hβ2m monomers when cross-seeded with fibrils formed from the N-terminally truncated variant ΔN6-hβ2m. Together, these results provide detailed insights into how collagen I and LMW-heparin impact different stages in the aggregation of WT-hβ2m into amyloid which lead to dramatic effects on the time course of assembly.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2019 Benseny-Cases et al. Published by The American Society for Biochemistry and Molecular Biology, Inc. This is an open access article under the terms of the Creative Commons: CC-BY license. |
Keywords: | β2-microglobulin; heparin; dilaysis ralted amyloidosis (DRA); MHC I; glycosaminoglycan; amyloid; collagen; protein aggregation; fibril; protein misfolding; extracellular matrix |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 01 May 2019 14:31 |
Last Modified: | 28 Jun 2019 14:26 |
Status: | Published |
Publisher: | American Society for Biochemistry and Molecular Biology |
Identification Number: | 10.1074/jbc.ra119.008300 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:145479 |