Skip to main content
U.S. Department of Energy
Office of Scientific and Technical Information

Purification of a major Cu-binding protein from mouse hepatic cytosols

Conference · · FASEB Journal (Federation of American Societies for Experimental Biology); (United States)
OSTI ID:5373063
;  [1]
  1. State Univ. of New York, Buffalo (United States)

Cytosolic Cu-binding proteins were purified by HPLC and SDS-PAGE. The goal was to identify proteins which may be involved in intracellular copper metabolism. When {sup 64}Cu-labeled cytosols were applied to Superose columns, two major {sup 64}Cu-binding fractions were detected which peaked at 130-kDa (I{sub s}) and 50-kDa (II{sub s}). These proteins were partially purified by successive Mono Q, re-chromatography on Mono Q and Superose steps. When one specific, {sup 64}Cu-labeled Mono Q fraction was applied to Superose, three {sup 64}Cu-binding fractions were detected. Two of these eluted near the I{sub s} peak and one near the II{sub s} peak. SDS-PAGE suggested that each of these Superose fractions contained the same 50-kDa subunit which was responsible for the Cu-binding. The 50-kDa protein was purified to homogeneity by sequential Mono Q, Chelating Superose, Phenyl Superose and Superose steps. The purified protein apparently exists as a monomer-dimer-tetramer equilibrium. The tetramer and dimer elute within the I{sub s} fraction, and the monomer elutes in the II{sub s} fraction. The same protein from the brindled mouse model of Menkes disease appeared to have a decreased affinity for copper. This protein is likely to play an important role in intracellular copper-trafficking.

OSTI ID:
5373063
Report Number(s):
CONF-9104107--
Journal Information:
FASEB Journal (Federation of American Societies for Experimental Biology); (United States), Journal Name: FASEB Journal (Federation of American Societies for Experimental Biology); (United States) Vol. 5:4; ISSN 0892-6638; ISSN FAJOE
Country of Publication:
United States
Language:
English