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Purification and functional reconstitution of the human Wilson copper ATPase, ATP7B
Author(s) -
Portmann Reto,
Solioz Marc
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.05.042
Subject(s) - atpase , affinity chromatography , biochemistry , histidine , copper , enzyme , yeast , chemistry , wilson's disease , biology , disease , organic chemistry , medicine , pathology
Wilson disease is a disorder of copper metabolism, due to inherited mutations in the Wilson copper ATPase gene ATP7B . To purify and study the function of the ATPase, the enzyme was truncated by five of the six metal binding domains and endowed with an N‐terminal histidine‐tag for affinity purification. This construct, Δ1–5WNDP, was able to functionally complement a yeast strain defective in its native copper ATPase CCC2. Δ1–5WNDP was purified by Ni‐affinity chromatography and reconstituted into proteoliposomes. This allowed, for the first time, the functional study of the Wilson ATPase in a purified, reconstituted system.