The cellular processes which regulate the uptake, storage, utilization, and excretion of the essential trace element, copper, are to be established. Incorporation of copper within liver cells into the serum protein ceruloplasmin will be central to our approach. Thus, while elucidating exactly how copper reaches apoceruloplasmin and is then incorporated, most aspects of liver cell copper metabolism must be examined. Both in vitro and in vivo experiments will be carried out using recently developed techniques of immunological electron and fluorescence microscopy and subcellular fractionation. The compartmentalization of apoprotein synthesis and available metal ion will be assessed as a mechanism for specificity of metal incorporation. In vitro metal binding to apoproteins will be studied to locate any factor(s) in the separate cellular fractions which may facilitate the rate and/or specificity of incorporation. To examine metalloprotein-biosynthesis under conditions which are more amenable to regulating levels of copper and each related metal, copper incorporation into galactose oxidase, a secretory protein of the eucaryotic fungus, P. circinatus, will also be carefully examined. Relationships between normal copper metabolism in liver and Wilson's Disease will be carefully considered. Moreover, the physiological, biochemical and pathological characteristics of a liver disease which is characterized by elevated liver copper levels in Bedlington terriers will be carefully evaluated as an animal model for Wilson's Disease.