Functional analysis of the sheep Wilson disease protein (sATP7B) in CHO cells

Lockhart, Paul J. and Mercer, Julian 2001, Functional analysis of the sheep Wilson disease protein (sATP7B) in CHO cells, European journal of cell biology, vol. 80, no. 5, pp. 349-357, doi: 10.1078/0171-9335-00165.

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Title Functional analysis of the sheep Wilson disease protein (sATP7B) in CHO cells
Author(s) Lockhart, Paul J.
Mercer, Julian
Journal name European journal of cell biology
Volume number 80
Issue number 5
Start page 349
End page 357
Publisher Elsevier GmbH
Place of publication Amsterdam, The Netherlands
Publication date 2001-05
ISSN 0171-9335
Keyword(s) copper transport
P-type ATPase
Summary In this study we investigated the function of the sheep orthologue of ATP7B (sATP7B), the protein affected in the human copper toxicosis disorder Wilson disease. Two forms of sATP7B are found in the sheep, a ‘normal’ form and one with an alternate N terminus, both of which were expressed in CHO-K1 cells. Cells expressing either form of sATP7B were more resistant to copper than the parental CHO-K1 cells. Subcellular localisation studies showed that both forms of sATP7B were similarly located in the trans-Golgi network (TGN). When the extracellular copper concentration was increased, each form of sATP7B redistributed to a punctate, vesicular compartment that extended throughout the cytoplasm. Both forms of sATP7B recycled to the perinuclear location within one hour when the cells were subsequently incubated in basal medium. After treatment of cells with bafilomycin A1 sATP7B accumulated in cytoplasmic vesicles, implying that ATP7B continuously recycles via the endocytic pathway. These results suggest that both forms of sATP7B are functional copper-transport proteins and that the intracellular location and trafficking of the sheep protein within the cell also appears normal.
Language eng
DOI 10.1078/0171-9335-00165
Field of Research 060199 Biochemistry and Cell Biology not elsewhere classified
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2001 Urban & Fischer Verlag
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