Partial purification, characterization and translation in vitro of rat liver metallothionein messenger ribonucleic acid
| Autor: | R D Andersen, U Weser |
|---|---|
| Rok vydání: | 1978 |
| Předmět: |
inorganic chemicals
In Vitro Techniques Biology Biochemistry Ribosome Serine chemistry.chemical_compound Biosynthesis Polysome Metalloproteins Centrifugation Density Gradient Animals Metallothionein RNA Messenger Molecular Biology Messenger RNA Translation (biology) Cell Biology Molecular biology Rats Kinetics Zinc Liver chemistry Polyribosomes Protein Biosynthesis Electrophoresis Polyacrylamide Gel Female Poly A Cadmium Research Article Cysteine |
| Zdroj: | Biochemical Journal. 175:841-852 |
| ISSN: | 0264-6021 |
| DOI: | 10.1042/bj1750841 |
| Popis: | Poly(A)+ (polyadenylated) mRNA coding for metallothioneins was purified 13-fold from rat liver polyribosomes and was identified by its ability to direct the biosynthesis of these proteins in a wheat-germ cell-free system. The carboxymethylated products of the protein-synthesizing system in vitro were analysed with sodium dodecyl sulphate/20% polyacrylamide-gel electrophoresis. The labelled compounds [3H]serine and [35S]cysteine were incorporated at high specific radioactivity into proteins that co-migrated with authentic metallothioneins. No [3H]leucine incorporation was found, in agreement with the amino acid composition of the metallothioneins. Metallothionein mRNA had a sedimentation coefficient of 9 S and carried a maximum of four ribosomes. At 5 h after a subcutaneous injection of ZnCl2 or CdCl2 (10 mumol/kg body wt.), the amount of this mRNA increased approx. 2- and 4-fold respectively, on the basis of translation in vitro. The increase in metallothionein mRNA (defined by translation in the wheat-germ system) was transient and, after CdCl2 treatment, fell back to control values by 17 h. Metallothioneins constituted a maximum of 0.8% of the total protein products synthesized in the wheat-germ system by total mRNA isolated from rat liver after CdCl2 treatment. |
| Databáze: | OpenAIRE |
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