Induction of Human High KM 5′-Nucleotidase in Cultured 293 Cells
| Autor: | Vera Bianchi, Monica Moras, Peter Reichard, Cinzia Gazziola, Paola Ferraro, Roberta Verin, Lisa Gallinaro, Chiara Rampazzo |
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| Rok vydání: | 1999 |
| Předmět: |
Receptors
Steroid DNA Complementary Nucleotidase activity Cytidine Triphosphate Protein subunit Blotting Western Green Fluorescent Proteins Antineoplastic Agents Uridine Triphosphate Biology Kidney Transfection Gene Expression Regulation Enzymologic Green fluorescent protein 5'-nucleotidase Adenosine Triphosphate Genes Reporter Nucleotidase Complementary DNA Humans Cloning Molecular 5'-Nucleotidase Cells Cultured Expression vector Cell Biology Molecular biology Enzyme Activation Luminescent Proteins Ecdysterone Biochemistry Cladribine Indicators and Reagents Guanosine Triphosphate Plasmids |
| Zdroj: | Experimental Cell Research. 253:474-482 |
| ISSN: | 0014-4827 |
| DOI: | 10.1006/excr.1999.4681 |
| Popis: | Human 293 cells were stably transfected with a plasmid introducing a receptor for the ecdysone analog muristerone. The cells were further stably transfected with muristerone-inducible expression vectors carrying either the cDNA for the human high K(M) 5'-nucleotidase or the coding sequence of the nucleotidase linked to the 5'-end of the sequence for the green fluorescent protein. Upon induction, both types of transfectants overproduced nucleotidase activity in a time- and dose-dependent manner. Western blots gave values close to the expected subunit molecular masses of 65 and 92 kDa, respectively, excluding processing of the induced proteins. Cells induced to overexpress the nucleotidase showed a decreased growth rate and contained smaller pools of each of the four common ribonucleoside triphosphates. They showed no increased resistance to the toxicity of 2-chlorodeoxyadenosine. |
| Databáze: | OpenAIRE |
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