2614

THE STUDY OF COPPER INDUCED ATPASE ACTIVITY FOR INBORN ERROR OF COPPER METABOLISM

Watanabe A1), Uno K1), Shimizu N1), Hemmi H2), Shimatake H2), Aoki T1)

1) 2nd Department of Pediatrics, Toho University School of Medicine, Ohashi Hospital

2) Department of Molecular Biology, Toho University School of Medicine, Japan

Introduction: Wilson disease and Menkes disease are inherited disorders of copper metabolism. The genes for these diseases are predicted to encode P-type ATPase involved in the transport of copper across membrane. We measured copper induced ATPase activities for these disorders.

Materials and Methods: The lymphoblastoid cell lines that delivered from one case of classical Menkes disease patient and his mother, five Japanese patients with Wilson disease, and two normal subjects were investigated. The non-copper stimulated ATPase activities of crude membranes were inhibited by sodium azide, ouabain and zinc sulfate. Then theses mixures were incubated at 37℃ for 60 min with copper sulfate and ATP, and copper-induced ATPase activities were calculated by measurement of released inorganic phosphate from ATP.

Results: Copper-induced ATPase activities were measured relative to the lymphoblast cell lines derived from normal and affected individuals. Loss of copper-induced ATPase activities in lymphoblast were 29.1±4.2%(Menkes disease patient), 48.2±1.9% (his mother) and 42.7-60.8% (Wilson disease patients).

Discussion: In this study, copper-induced ATPase activities of Menkes disease patient, carrier of this disease and Wilson disease patients were definitely lower than that of normal subjects. This method will be useful as a functional analysis for copper metabolic disorders.