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Effects of seeding on lysozyme amyloid fibrillation in the presence of epigallocatechin and polyethylene glycol

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Abstract

Preformed amyloid fibrils can act as seeds for accelerating protein fibrillation. In the present study, we examined the effects of preformed seeds on lysozyme amyloid fibrillation in the presence of two distinct inhibitors–epigallocatechin (EGC) and polyethylene glycol 2000 (PEG). The results demonstrated that the effects of fibrillar seeds on the acceleration of lysozyme fibrillation depended on the aggregation pathway directed by an inhibitor. EGC inhibited lysozyme fibrillation and modified the peptide chains with quinone moieties in a concentration-dependent manner. The resulting aggregates showed amorphous off-pathway morphology. Preformed fibril seeds did not promote lysozyme fibrillation in the presence of EGC. PEG also inhibited lysozyme fibrillation, and the resulting aggregates showed on-pathway protofibrillar morphology. In contrast, the addition of fibril seeds into the mixture of lysozyme and PEG significantly stimulated fibril growth. Assays of cell viability showed that both EGC and PEG inhibited the formation of cytotoxic species. In accordance with thioflavine T data, the seeds failed to alter the cell-damaging potency of the EGC-directed off-pathway aggregates, but increased the cytotoxicity of the PEG-directed on-pathway fibrils. We suggest that the pattern of interaction between lysozyme and an inhibitor determines the pathway of aggregation and therefore the effects of seeding on amyloid formation. EGC covalently modified lysozyme chains with quinones, directing the aggregation to proceed through an off-pathway, whereas PEG affected the protein in a noncovalent manner, and fibril growth could be stimulated under seeding through an on-pathway.

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Abbreviations

EGC:

epigallocatechin

EGCG:

epigallocatechin-3-gallate

MTT:

thiazolyl blue tetrazolium bromide

NBT:

nitroblue tetrazolium

PEG:

polyethylene glycol

TEM:

trans-mission electron microscopy

ThT:

thioflavine T

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  1. Shaanxi Normal University, School of Chemistry and Chemical Engineering, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, 710119, Xi’an, China

    Li-Xiu Kong & Cheng-Ming Zeng

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  1. Li-Xiu Kong
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Correspondence to Cheng-Ming Zeng.

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Published in Russian in Biokhimiya, 2017, Vol. 82, No. 2, pp. 266-279.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM16-252, November 7, 2016.

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Kong, LX., Zeng, CM. Effects of seeding on lysozyme amyloid fibrillation in the presence of epigallocatechin and polyethylene glycol. Biochemistry Moscow 82, 156–167 (2017). https://doi.org/10.1134/S0006297917020079

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