The rapid isolation of highly purified cadmium metallothioneins using HPLC

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

OSTI ID:5863395

Vella, G; Bylander, J; Hazen-martin, D; O'Connor, K; Phoebe, C ^[1]

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1. Univ. of South Carolina, Charleston (United States) Millipore Corp., Milford, MA (United States)

The central role of the proximal tubule in the renal toxicity due to cadmium (Cd) poisoning has been well documented, however, the mechanisms leading to the onset of this nephrotoxic effect are presently not well understood. The most likely mechanism for the initiation of toxicity involves the accumulation of Cd in the liver where it induces the formation of, and is complexed with the metallothioneins (MT-I and MT-II) followed by release from the liver and subsequent absorption by the proximal tubule. Methods for obtaining MTs for specific RIA and immunohistochemistry have been hampered by difficulties in removing impurities. To obtain highly purified Cd-MTs for use in RIA as well as to assay the relative affects of exposure to Cd-MT and ionic Cd in vitro on human proximal tubule cells, Cd-MT-I and Cd-MT-II were prepared from rat liver for use in these studies using a rapid two step chromatographic procedure employing gel filtration and high performance anion exchange chromatography. Repeated HPLC separations revealed a time dependent conversion of a non-metallothionein protein component to form which coeluted with Cd-MT-I during the ion exchange separation. This was preventable by cooling the sample or by adding proteinase inhibitors. This method demonstrated the importance of speed in the purification which permitted the isolation of highly purified Cd-MTs in high yields. Due to the extreme sensitivity, this method may also be amenable as an alternative for a non-radioactive assay of Cd-MT formation and breakdown in toxicological studies.

• OSTI ID: 5863395
• Report Number(s): CONF-9104107-; CODEN: FAJOE
• Journal Information: FASEB Journal (Federation of American Societies for Experimental Biology); (United States), Vol. 5:4; Conference: 75. annual meeting of the Federation of American Societies for Experimental Biology (FASEB), Atlanta, GA (United States), 21-25 Apr 1991; ISSN 0892-6638
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
CADMIUM
TOXICITY
METALLOTHIONEIN
FRACTIONATION
LIQUID COLUMN CHROMATOGRAPHY
LIVER
METABOLISM
RADIOIMMUNOASSAY
RATS
TIME DEPENDENCE
TUBULES
ANIMALS
BIOASSAY
BODY
CHROMATOGRAPHY
DIAGNOSTIC TECHNIQUES
DIGESTIVE SYSTEM
ELEMENTS
GLANDS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
KIDNEYS
MAMMALS
METALLOPROTEINS
METALS
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
RODENTS
SEPARATION PROCESSES
TRACER TECHNIQUES
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology