A mercury saturation assay for measuring metallothionein in fish
- Univ. of Manitoba, Winnipeg (Canada). Dept. of Zoology
- AECL Research, Pinawa, Manitoba (Canada). Environmental Science Branch
- Freshwater Inst., Winnipeg, Manitoba (Canada)
An accurate, rapid, sensitive, and simple method using mercury saturation for quantifying metallothionein (MT) is described. A complex solution of enzymatic and nonenzymatic thiols, including rabbit liver MT-2, and supernatants from homogenized samples of rainbow trout liver were incubated in the presence of [sup 203]Hg in 10% trichloroacetic acid. Excess Hg was bound to an removed by chicken egg albumin, which denatured on contact with the acidic assay medium. After centrifugation, MT labeled with [sup 203]Hg remained in the TCA supernatant and was estimated using known stoichiometry for Hg-MT binding. A dilution series was used to establish that nonspecific metal binding, a common problem with other metal saturation assays, is negligible. Analysis of hepatic MT with high Cu content from rainbow trout demonstrated virtually complete displacement of Cu, Cd, and Zn by Hg. When compared to other metal-saturation assays developed for vertebrates, this method requires the least number of technical steps, and one-third or less of total preparatory and analytical time.
- OSTI ID:
- 5592413
- Journal Information:
- Environmental Toxicology and Chemistry; (United States), Vol. 12:7; ISSN 0730-7268
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CADMIUM
BIOASSAY
COPPER
MERCURY
METALLOTHIONEIN
BIOSYNTHESIS
TROUT
SENSITIVITY
ZINC
METABOLISM
ANIMALS
AQUATIC ORGANISMS
ELEMENTS
FISHES
METALLOPROTEINS
METALS
ORGANIC COMPOUNDS
PROTEINS
SYNTHESIS
TRANSITION ELEMENTS
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology