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Enhanced molecular chaperone activity of the small heat-shock protein alphaB-cystallin following covalent immobilization onto a solid-phase support

Journal Article


Abstract


  • The well-characterized small heat-shock protein, alphaB-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 alphaB-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 alphaB-crystallin when covalently bound to a support surface; alphaB-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 alphaB-crystallin lysine residues to the surface aldehyde groups via Schiff-base linkages. Immobilized alphaB-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 alphaB-crystallin was bound to the surface. Immobilized alphaB-crystallin exhibited a significant enhancement (up to 5000-fold, when compared with the equivalent activity of alphaB-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 alphaB-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.

Authors


  •   Garvey, J (external author)
  •   Griesser, Stefani S. (external author)
  •   Griesser, Hans J. (external author)
  •   Thierry, B (external author)
  •   Nussio, M R (external author)
  •   Shapter, Joe G. (external author)
  •   Ecroyd, Heath
  •   Giorgetti, S (external author)
  •   Bellotti, V (external author)
  •   Gerrard, Juliet A. (external author)
  •   Carver, John A. (external author)

Publication Date


  • 2011

Citation


  • Garvey, M., Griesser, S. S., Griesser, H. J., Thierry, B., Nussio, M. R., Shapter, J. E., Ecroyd, H., Giorgetti, S., Bellotti, V., Gerrard, J. A. & Carver, J. A. (2011). Enhanced molecular chaperone activity of the small heat-shock protein alphaB-cystallin following covalent immobilization onto a solid-phase support. Biopolymers, 95 (6), 376-389.

Scopus Eid


  • 2-s2.0-80051740888

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/4101

Has Global Citation Frequency


Number Of Pages


  • 13

Start Page


  • 376

End Page


  • 389

Volume


  • 95

Issue


  • 6

Abstract


  • The well-characterized small heat-shock protein, alphaB-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 alphaB-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 alphaB-crystallin when covalently bound to a support surface; alphaB-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 alphaB-crystallin lysine residues to the surface aldehyde groups via Schiff-base linkages. Immobilized alphaB-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 alphaB-crystallin was bound to the surface. Immobilized alphaB-crystallin exhibited a significant enhancement (up to 5000-fold, when compared with the equivalent activity of alphaB-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 alphaB-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.

Authors


  •   Garvey, J (external author)
  •   Griesser, Stefani S. (external author)
  •   Griesser, Hans J. (external author)
  •   Thierry, B (external author)
  •   Nussio, M R (external author)
  •   Shapter, Joe G. (external author)
  •   Ecroyd, Heath
  •   Giorgetti, S (external author)
  •   Bellotti, V (external author)
  •   Gerrard, Juliet A. (external author)
  •   Carver, John A. (external author)

Publication Date


  • 2011

Citation


  • Garvey, M., Griesser, S. S., Griesser, H. J., Thierry, B., Nussio, M. R., Shapter, J. E., Ecroyd, H., Giorgetti, S., Bellotti, V., Gerrard, J. A. & Carver, J. A. (2011). Enhanced molecular chaperone activity of the small heat-shock protein alphaB-cystallin following covalent immobilization onto a solid-phase support. Biopolymers, 95 (6), 376-389.

Scopus Eid


  • 2-s2.0-80051740888

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/4101

Has Global Citation Frequency


Number Of Pages


  • 13

Start Page


  • 376

End Page


  • 389

Volume


  • 95

Issue


  • 6