Oligomers, protofibrils and amyloid fibrils from recombinant human lysozyme (rHL): Fibrillation process and cytotoxicity evaluation for ARPE-19 cell line

Eva D. Ruiz, Mario Almada, María G. Burboa, Pablo Taboada, Víctor Mosquera, Miguel A. Valdez, Josué Juárez*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Amyloid-associated diseases, such Alzheimer's, Huntington's, Parkinson's, and type II diabetes, are related to protein misfolding and aggregation. Herein, the time evolution of scattered light intensity, hydrophobic properties, and conformational changes during fibrillation processes of rHL solutions at 55. °C and pH 2.0 were used to monitor the aggregation process of recombinant human lysozyme (rHL). Dynamic light scattering (DLS), thioflavin T (ThT) fluorescence, and surface tension (ST) at the air-water interface were used to analyze the hydrophobic properties of pre-amyloid aggregates involved in the fibrillation process of rHL to find a correlation between the hydrophobic character of oligomers, protofibrils and amyloid aggregates with the gain in cross-β-sheet structure, depending on the increase in the incubation periods. The ability of the different aggregates of rHL isolated during the fibrillation process to be adsorbed at the air-water interface can provide important information about the hydrophobic properties of the protein, which can be related to changes in the secondary structure of rHL, resulting in cytotoxic or non-cytotoxic species. Thus, we evaluated the cytotoxic effect of oligomers, protofibrils and amyloid fibrils on the cell line ARPE-19 using the MTT reduction test. The more cytotoxic protein species arose after a 600-min incubation time, suggesting that the hydrophobic character of pre-amyloid fibrils, in addition to the high prevalence of the cross-β-sheet conformation, can become toxic for the cell line ARPE-19.

Original languageEnglish
Pages (from-to)335-343
Number of pages9
JournalColloids and Surfaces B: Biointerfaces
Volume126
DOIs
StatePublished - 1 Feb 2015

Bibliographical note

Publisher Copyright:
© 2014 Elsevier B.V.

Keywords

  • ARPE-19
  • Amyloid fibrils
  • Oligomers
  • Protofibrils
  • Recombinant human lysozyme

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