Stabilization, Characterization and Selective Removal of Cystatin C Amyloid Oligomers.

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

The pathophysiological process in amyloid disorders usually involves the transformation of a functional monomeric protein via potentially toxic oligomers into amyloid fibrils. The structure and properties of the intermediary oligomers have been difficult to study due to their instability and dynamic equilibrium with smaller and larger species. In hereditary cystatin C amyloid angiopathy, a cystatin C variant is deposited in arterial walls and cause brain hemorrhage in young adults. In the present investigation, we use redox experiments of monomeric cystatin C, stabilized against domain swapping by an intramolecular disulfide bond, to generate stable oligomers (dimers, trimers, tetramers, decamers and high molecular weight oligomers). These oligomers were characterized concerning size by gel filtration, polyacrylamide gel electrophoresis and mass spectrometry, concerning shape by electron and atomic force microscopy, and, concerning function, by assays of their capacity to inhibit proteases. The results showed the oligomers to be highly ordered, domainswapped assemblies of cystatin C and that the oligomers could not build larger oligomers, or fibrils, without domain swapping. The stabilized oligomers were used to induce antibody formation in rabbits. After immunosorption, using immobilized monomeric cystatin C, and elution from columns with immobilized cystatin C oligomers, oligomer-specific antibodies were obtained. These could be used to selectively remove cystatin C dimers from biological fluids containing both dimers and monomers.

Detaljer

Författare
Enheter & grupper
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Farmakologi och toxikologi
  • Klinisk laboratoriemedicin
Originalspråkengelska
Sidor (från-till)16438-16450
TidskriftJournal of Biological Chemistry
Volym288
Utgåva nummer23
StatusPublished - 2013
PublikationskategoriForskning
Peer review utfördJa

Nedladdningar

Ingen tillgänglig data

Relaterad forskningsoutput

Gustav Östner, 2013, Division of Clinical Chemistry and Pharmacology, Faculty of Medicine, Lund University. 98 s.

Forskningsoutput: AvhandlingDoktorsavhandling (sammanläggning)

Visa alla (1)