Enhanced molecular chaperone activity of the small heat-shock protein {alpha}B-crystallin following covalent immobilization onto a solid-phase support

Megan Garvey, Stefani Griesser, Hans Griesser, Benjamin Thierry, Matthew Nussio, Joseph Shapter, Heath Ecroyd, Sofia Giorgetti, Vittorio Bellotti, Juliet Gerrard, John Carver

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    Abstract

    The well-characterized small heat-shock protein, αB-crystallin, acts as a molecular chaperone by interacting with unfolding proteins to prevent their aggregation and precipitation. Structural perturbation (e.g., partial unfolding) enhances the in vitro chaperone activity of αB-crystallin. Proteins often undergo structural perturbations at the surface of a synthetic material, which may alter their biological activity. This study investigated the activity of αB-crystallin when covalently bound to a support surface; αB-crystallin was immobilized onto a range of solid material surfaces, and its characteristics and chaperone activity were assessed. Immobilization was achieved via a plasma-deposited thin polymeric interlayer containing aldehyde surface groups and reductive amination, leading to the covalent binding of αB-crystallin lysine residues to the surface aldehyde groups via Schiff-base linkages. Immobilized αB-crystallin was characterized by X-ray photoelectron spectroscopy, atomic force microscopy, and quartz crystal microgravimetry, which showed that ∼300 ng cm -2 (dry mass) of oligomeric αB-crystallin was bound to the surface. Immobilized αB-crystallin exhibited a significant enhancement (up to 5000-fold, when compared with the equivalent activity of αB-crystallin in solution) of its chaperone activity against various proteins undergoing both amorphous and amyloid fibril forms of aggregation. The enhanced molecular chaperone activity of immobilized αB-crystallin has potential applications in preventing protein misfolding, including against amyloid disease processes, such as dialysis-related amyloidosis, and for biodiagnostic detection of misfolded proteins.

    Original languageEnglish
    Pages (from-to)376-389
    Number of pages14
    JournalBiopolymers
    Volume95
    Issue number6
    DOIs
    Publication statusPublished - 2011

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  • Cite this

    Garvey, M., Griesser, S., Griesser, H., Thierry, B., Nussio, M., Shapter, J., Ecroyd, H., Giorgetti, S., Bellotti, V., Gerrard, J., & Carver, J. (2011). Enhanced molecular chaperone activity of the small heat-shock protein {alpha}B-crystallin following covalent immobilization onto a solid-phase support. Biopolymers, 95(6), 376-389. https://doi.org/10.1002/bip.21584