Using XAS and SXRF to study copper in wilson disease at the molecular and tissue level

Martina Ralle, Ninian Blackburn, Svetlana Lutsenko

Research output: Chapter in Book/Report/Conference proceedingConference contribution

6 Citations (Scopus)

Abstract

Wilson disease (WD) is a genetic disorder of copper metabolism associated with severe hepatic, neurological, and psychiatric abnormalities. In WD, the billiary copper excretion is impaired and copper accumulates in tissues, particularly in the liver and the brain. The affected gene, ATP7B, encodes the copper transporting ATPase, Wilson disease protein (WNDP). WNDP has six copper binding sites in the N-terminal portion of the molecule. Each site includes the conserved amino acid sequence MXCXXC, and binds 1 Cu(I) through its 2 cysteine residues. We performed X-ray absorption studies at the Cu Kα-edge on the recombinant N-terminal domain of WNDP (N-WNDP). Copper was bound to N-WNDP either in vivo or in vitro in the presence of different reducing agents. We found that in N-WNDP copper is predominantly coordinated in a linear fashion by two cysteines, with the appearance of a Cu-Cu interaction when all metal binding sites are filled. Increasing amounts of reducing agents containing sulfide or phosphine groups led to binding of the exogenous ligands to copper thereby increasing the coordination number of copper from two to three. To better understand the role of copper in WD, we utilized livers of the 6-weeks-old Atp7b-/- mice (an animal model for WD) in which the copper concentration was 10-20-fold higher compared to that of the control mice. The distribution of copper in hepatocytes was evaluated by synchrotron based X-ray fluorescence microprobe (SXRF). We demonstrate that we can prepare liver slices that retain copper and can detect copper with subcellular resolution. On the same sections μ-XANES (spot size: 5 micron) was used to determine the oxidation state of copper.

Original languageEnglish (US)
Title of host publicationAIP Conference Proceedings
Pages328-330
Number of pages3
Volume882
DOIs
StatePublished - 2007
EventX-RAY ABSORPTION FINE STRUCTURE - XAFS13: 13th International Conference - Stanford, CA, United States
Duration: Jul 9 2006Jul 14 2006

Other

OtherX-RAY ABSORPTION FINE STRUCTURE - XAFS13: 13th International Conference
CountryUnited States
CityStanford, CA
Period7/9/067/14/06

Fingerprint

synchrotrons
copper
fluorescence
x rays
liver
cysteine
proteins
mice
excretion
animal models
abnormalities
metabolism
coordination number
phosphines
genes
brain
amino acids
sulfides
disorders

Keywords

  • Copper
  • EXAFS
  • SXRF
  • Wilson disease

ASJC Scopus subject areas

  • Physics and Astronomy(all)

Cite this

Using XAS and SXRF to study copper in wilson disease at the molecular and tissue level. / Ralle, Martina; Blackburn, Ninian; Lutsenko, Svetlana.

AIP Conference Proceedings. Vol. 882 2007. p. 328-330.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Ralle, M, Blackburn, N & Lutsenko, S 2007, Using XAS and SXRF to study copper in wilson disease at the molecular and tissue level. in AIP Conference Proceedings. vol. 882, pp. 328-330, X-RAY ABSORPTION FINE STRUCTURE - XAFS13: 13th International Conference, Stanford, CA, United States, 7/9/06. https://doi.org/10.1063/1.2644515
Ralle, Martina ; Blackburn, Ninian ; Lutsenko, Svetlana. / Using XAS and SXRF to study copper in wilson disease at the molecular and tissue level. AIP Conference Proceedings. Vol. 882 2007. pp. 328-330
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