Replacement of All α-Domain Lysines with Glutamates Reduces Metallothionein Detoxification Function
| Autor: | P. C. Huang, Chris W. Cody |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Mutant
Lysine Drug Resistance Biophysics Glutamic Acid Mutagenesis (molecular biology technique) Saccharomyces cerevisiae Biology complex mixtures Biochemistry Structure-Activity Relationship Glutamates Metallothionein Molecular Biology Binding Sites Chinese hamster ovary cell Gene Transfer Techniques Wild type Cell Biology Glutamic acid Yeast Mutagenesis Site-Directed bacteria Electrophoresis Polyacrylamide Gel Cadmium |
| Zdroj: | Biochemical and Biophysical Research Communications. 202:954-959 |
| ISSN: | 0006-291X |
| DOI: | 10.1006/bbrc.1994.2022 |
| Popis: | Mammalian metallothioneins (MTs) possess eight highly conserved lysine residues, including three in each of two metal binding domains. We used site-directed mutagenesis to replace these intradomain lysines in Chinese hamster ovary MT2 with glutamic acid and/or glutamine. These mutant MTs were expressed in a metal sensitive yeast host. One mutant which had all three lysines in the α-domain replaced by glutamates (K43,51,56E) exhibited a reduced ability, relative to native MT, to protect yeast transformants against otherwise toxic levels of cadmium. This triply substituted mutant also exhibited anomalous migration on a non-denaturing gel relative to wild type MT and other MT lysine mutants, suggesting that the intradomain lysines are important in maintaining the conformational integrity of MT. |
| Databáze: | OpenAIRE |
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