Mimicking kidney flow shear efficiently induces aggregation of LECT2, a protein involved in renal amyloidosis

Ha, Jeung-Hoi; Xu, Yikang; Sekhon, Harsimranjit; Zhao, Wenhan; Wilkens, Stephan; Ren, Dacheng; Loh, Stewart N.

doi:10.1016/j.jbc.2024.107231

Cited by 2 publications

References 62 publications

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The role of shear forces in primary and secondary nucleation of amyloid fibrils

Axell,

Hu,

Lindberg

et al. 2024

Proc. Natl. Acad. Sci. U.S.A.

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Shear forces affect self-assembly processes ranging from crystallization to fiber formation. Here, the effect of mild agitation on amyloid fibril formation was explored for four peptides and investigated in detail for A β 42, which is associated with Alzheimer’s disease. To gain mechanistic insights into the effect of mild agitation, nonseeded and seeded aggregation reactions were set up at various peptide concentrations with and without an inhibitor. First, an effect on fibril fragmentation was excluded by comparing the monomer-concentration dependence of aggregation kinetics under idle and agitated conditions. Second, using a secondary nucleation inhibitor, Brichos, the agitation effect on primary nucleation was decoupled from secondary nucleation. Third, an effect on secondary nucleation was established in the absence of inhibitor. Fourth, an effect on elongation was excluded by comparing the seeding potency of fibrils formed under idle or agitated conditions. We find that both primary and secondary nucleation steps are accelerated by gentle agitation. The increased shear forces facilitate both the detachment of newly formed aggregates from catalytic surfaces and the rate at which molecules are transported in the bulk solution to encounter nucleation sites on the fibril and other surfaces. Ultrastructural evidence obtained with cryogenic transmission electron microscopy and free-flow electrophoresis in microfluidics devices imply that agitation speeds up the detachment of nucleated species from the fibril surface. Our findings shed light on the aggregation mechanism and the role of detachment for efficient secondary nucleation. The results inform on how to modulate the relative importance of different microscopic steps in drug discovery and investigations.

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The role of shear forces in primary and secondary nucleation of amyloid fibrils

Axell,

Hu,

Lindberg

et al. 2024

Proc. Natl. Acad. Sci. U.S.A.

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Peristaltic pump-triggered amyloid formation suggests shear stresses arein vivorisk factors of amyloid nucleation

Goto,

Ota,

Yuan

et al. 2024

Preprint

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Amyloid fibrils, crystal-like fibrillar aggregates of denatured proteins, are formed linked with the breakdown of supersaturation, causing a series of amyloidosis including Alzheimer's and Parkinson's diseases. Although varying in vitro factors are known, in vivo factors breaking supersaturation are unclear. We found that flowing by a peristaltic pump effectively triggers amyloid formation of hen egg white lysozyme, a model amyloidogenic protein, and, moreover, amyloidosis-associated proteins (i.e. α-synuclein, amyloid β 1-40, and β2-microglobulin). The peristaltic pump-dependent amyloid formation was visualized by a fluorescence microscope with looped flow system, revealing dynamic motions under flow. Among them, amyloid fibrils of amyloid β 1-40 were stickier than others, self-associating, absorbing to loop surfaces, and surging upon flicking the loop, implying early stages of cerebral amyloid angiopathy. On the other hand, β2-microglobulin at a neutral pH showed unique two-step amyloid formation with an oligomeric trapped intermediate, which might mimic amyloid formation in patients. Peristalsis-caused strong shear stresses were considered to mechanically break supersaturation. Shearing stresses occur in vivo at varying levels, suggesting that they break otherwise persistent supersaturation, thus triggering amyloid formation and ultimately leading to amyloidosis.

show abstract

Mimicking kidney flow shear efficiently induces aggregation of LECT2, a protein involved in renal amyloidosis

Cited by 2 publications

References 62 publications

The role of shear forces in primary and secondary nucleation of amyloid fibrils

The role of shear forces in primary and secondary nucleation of amyloid fibrils

Peristaltic pump-triggered amyloid formation suggests shear stresses arein vivorisk factors of amyloid nucleation

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