Solid-phase detection of low molecular weight zinc- and cadmium-binding proteins from rat intestine

Conference · Fri Mar 15 04:00:00 EST 1991 · FASEB Journal (Federation of American Societies for Experimental Biology); (United States)

OSTI ID:5239895

Hempe, J M; Cousins, R J ^[1]

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1. Univ. of Florida, Gainesville (United States)

Intracellular proteins from liver, pancreas, and intestinal mucosa were separated by Tricine SDS-PAGE or gel filtration HPLC, and transferred to nitrocellulose membranes by electroblotting or vacuum blotting, respectively. Following equilibration in blocking buffer the membranes were placed in buffer containing either {sup 65}Zn or {sup 109}Cd, and then washed in buffer without MnCl{sub 2} or radioisotope. Autoradiographs from SDS-PAGE blots showed little difference between control rats and those injected with ZnSO{sub 4}. Radioactive bands attributable to metallothionein were readily apparent in all three tissues of rats given ZnSO{sub 4} when reducing agent was added to the metal-binding buffers. Purified metallothionein showed multiple protein bands by SDS-PAGE, some of which bound radioisotope only in the presence of 2-mercaptoethanol, suggesting polymerization through sulfhydryl groups. Autoradiographs from both SDS-PAGE and HPLC showed the presence of a low-molecular-weight intestinal protein that is apparently not metallothionein and that readily binds {sup 65}Zn. These data corroborate in vivo labeling experiments, and showed that this zinc-binding protein is not present in either liver or pancreas, suggesting a possible role in zinc absorption.

• OSTI ID: 5239895
• Report Number(s): CONF-9104107--
• Conference Information: Journal Name: FASEB Journal (Federation of American Societies for Experimental Biology); (United States) Journal Volume: 5:5
• 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.
ABSORPTION
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-PLUS DECAY RADIOISOTOPES
BODY
CADMIUM
CADMIUM 109
CADMIUM ISOTOPES
CHEMICAL ANALYSIS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ELEMENTS
ENDOCRINE GLANDS
EVEN-ODD NUCLEI
GASTROINTESTINAL TRACT
GLANDS
INTERMEDIATE MASS NUCLEI
INTESTINAL ABSORPTION
INTESTINES
ISOTOPES
LIQUID COLUMN CHROMATOGRAPHY
LIVER
MAMMALS
MEMBRANES
METABOLISM
METALLOPROTEINS
METALLOTHIONEIN
METALS
MUCOUS MEMBRANES
NUCLEI
ORGANIC COMPOUNDS
ORGANS
PANCREAS
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SEPARATION PROCESSES
UPTAKE
VERTEBRATES
YEARS LIVING RADIOISOTOPES
ZINC
ZINC 65
ZINC ISOTOPES