Cloning and expression of cDNA encoding protein synthesis elongation factor-2 kinase
| Autor: | N T, Redpath, N T, Price, C G, Proud |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Elongation Factor 2 Kinase
DNA Complementary Reticulocytes Transcription Genetic Molecular Sequence Data Gene Expression Polymerase Chain Reaction Peptide Elongation Factor 2 Consensus Sequence Animals Amino Acid Sequence RNA Messenger Cloning Molecular Caenorhabditis elegans Muscle Skeletal Conserved Sequence DNA Primers Binding Sites Base Sequence Sequence Homology Amino Acid Peptide Elongation Factors Recombinant Proteins Rats Isoenzymes Molecular Weight Protein Biosynthesis Calcium-Calmodulin-Dependent Protein Kinases Rabbits |
| Zdroj: | The Journal of biological chemistry. 271(29) |
| ISSN: | 0021-9258 |
| Popis: | A cDNA from rat skeletal muscle encoding calcium/calmodulin-dependent eukaryotic elongation factor-2 kinase (eEF-2K) has been cloned and sequenced, and the amino acid sequence of the protein has been deduced. The kinase is composed of 724 amino acids and has a predicted molecular mass of 81,499 Da. The cDNA was judged to be full-length, as the protein, expressed in rabbit reticulocyte lysate or wheat germ extract, migrated upon SDS-PAGE with the same apparent molecular weight as the purified kinase and possessed eEF-2K activity. eEF-2K contains all of the 12 catalytic subdomains present in the majority of protein kinases, but they are atypical and display only limited homology with other kinases. A putative calmodulin-binding domain is present C-terminal to the catalytic domain as is a putative pseudosubstrate sequence. Two antipeptide antibodies raised against sequences derived from a partial rabbit cDNA clone, cross-reacted with purified eEF-2K, and one also immunoprecipitated eEF-2K activity from cell extracts. Northern blot analysis demonstrated that eEF-2K mRNA is expressed in a number of different tissues and that it may exist in multiple forms. |
| Databáze: | OpenAIRE |
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