Production of dog calcyphosine in bacteria and lack of phosphorylation by the catalytic subunit of protein kinase A in vitro
| Autor: | H El Housni, Daniel Christophe, Raymond Lecocq |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Protein subunit
Thyroid Gland Gene Expression Biochemistry law.invention Dogs Endocrinology law Complementary DNA Escherichia coli Animals Phosphorylation Protein kinase A Molecular Biology chemistry.chemical_classification biology Calcium-Binding Proteins biology.organism_classification Cyclic AMP-Dependent Protein Kinases Molecular biology Recombinant Proteins In vitro Enzyme chemistry Recombinant DNA Electrophoresis Polyacrylamide Gel Bacteria |
| Zdroj: | Molecular and Cellular Endocrinology. 135:93-97 |
| ISSN: | 0303-7207 |
| Popis: | Calcyphosine is a calcium-binding protein containing four EF-hand domains that is found in several epithelia and in some cells of the central nervous system. In thyroid follicular cells, calcyphosine is synthesized and phosphorylated in response to stimulation by thyrotropin and cAMP agonists. The cDNA coding for dog calcyphosine has been expressed in bacteria under the control of the T7 promoter. Recombinant calcyphosine was purified from crude bacterial lysates by a combination of anion-exchange and hydrophobic interaction chromatography. Phosphorylation assays using the purified catalytic subunit of protein kinase A and the recombinant or the native calcyphosine revealed that, contrary to a previous report, calcyphosine is not significantly phosphorylated by this enzyme in vitro. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect