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

Conference · Mon Mar 11 04:00:00 EST 1991 · FASEB Journal (Federation of American Societies for Experimental Biology); (United States)

OSTI ID:5373063

Seo, H C; Ettinger, M J ^[1]

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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--
• Conference Information: Journal Name: FASEB Journal (Federation of American Societies for Experimental Biology); (United States) Journal Volume: 5:4
• Country of Publication: United States
• Language: English

Related Subjects

560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BETA-PLUS DECAY RADIOISOTOPES
BODY
CHROMATOGRAPHY
COPPER
COPPER 64
COPPER ISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ELEMENTS
FRACTIONATION
GLANDS
HOURS LIVING RADIOISOTOPES
INTERMEDIATE MASS NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
LIQUID COLUMN CHROMATOGRAPHY
LIVER
MAMMALS
METABOLISM
METALLOPROTEINS
METALS
MICE
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOISOTOPES
RODENTS
SEPARATION PROCESSES
TRACER TECHNIQUES
TRANSITION ELEMENTS
VERTEBRATES