Quantitation of Cu-containing metallothionein by a Cd-saturation method

Journal Article · Wed Aug 15 04:00:00 EDT 1990 · Analytical Biochemistry; (USA)

DOI:https://doi.org/10.1016/0003-2697(90)90040-G· OSTI ID:5889000

Klein, D; Bartsch, R; Summer, K H ^[1]

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1. GSF Research Center, Neuherberg (Germany, F.R.)

A rapid and sensitive method for determining Cu-containing metallothionein (MT) is described. The main features of this Cd-saturation assay are: high-molecular-weight Cd-binding compounds are denatured with acetonitrile (50% final concentration), Cu bound to MT is removed with ammonium tetrathiomolybdate, excessive tetrathiomolybdate and its Cu complexes are removed with DEAE-Sephacel, apothionein is saturated with Cd, and excessive Cd is bound to Chelex 100. The thiomolybdate assay is capable of reliably detecting 14 ng MT and thus is particularly suitable for measuring MT in small tissue samples (e.g., biopsies), in extrahepatic tissues, and in cultured cells. Moreover, the combination of the thiomolybdate assay with the recently developed Cd-Chelex assay also makes it possible to determine the portion of MT which binds Cu (Cu load of MT), provided that the amount of non-Cu-thionein exceeds 100 ng, the detection limit of the Cd-Chelex assay.

• OSTI ID: 5889000
• Journal Information: Analytical Biochemistry; (USA), Journal Name: Analytical Biochemistry; (USA) Vol. 189:1; ISSN ANBCA; ISSN 0003-2697
• Country of Publication: United States
• Language: English

Related Subjects

550501 -- Metabolism-- Tracer Techniques
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
CADMIUM ISOTOPES
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
COPPER
DRUGS
ELEMENTS
FIBROBLASTS
GLUTATHIONE
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
MEASURING METHODS
METABOLISM
METALLOPROTEINS
METALLOTHIONEIN
METALS
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOPROTECTIVE SUBSTANCES
RATS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
TRACER TECHNIQUES
TRANSITION ELEMENTS
VERTEBRATES