Cloning, Expression in Escherichia coli, and Characterization of Arabidopsis thaliana UMP/CMP Kinase1
| العنوان: | Cloning, Expression in Escherichia coli, and Characterization of Arabidopsis thaliana UMP/CMP Kinase1 |
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| المؤلفون: | François Lacroute, Robert W. Thornburg, Lan Zhou |
| المصدر: | Plant Physiology. 117:245-254 |
| بيانات النشر: | Oxford University Press (OUP), 1998. |
| سنة النشر: | 1998 |
| مصطلحات موضوعية: | chemistry.chemical_classification, biology, Physiology, Kinase, Saccharomyces cerevisiae, Proteolytic enzymes, Plant Science, medicine.disease_cause, biology.organism_classification, Uridine, carbohydrates (lipids), chemistry.chemical_compound, Enzyme, chemistry, Biochemistry, Complementary DNA, Genetics, medicine, Escherichia coli, Peptide sequence |
| الوصف: | A cDNA encoding theArabidopsis thaliana uridine 5′-monophosphate (UMP)/cytidine 5′-monophosphate (CMP) kinase was isolated by complementation of a Saccharomyces cerevisiae ura6 mutant. The deduced amino acid sequence of the plant UMP/CMP kinase has 50% identity with other eukaryotic UMP/CMP kinase proteins. The cDNA was subcloned into pGEX-4T-3 and expressed as a glutathione S-transferase fusion protein in Escherichia coli. Following proteolytic digestion, the plant UMP/CMP kinase was purified and analyzed for its structural and kinetic properties. The mass, N-terminal sequence, and total amino acid composition agreed with the sequence and composition predicted from the cDNA sequence. Kinetic analysis revealed that the UMP/CMP kinase preferentially uses ATP (Michaelis constant [Km] = 29 μm when UMP is the other substrate andKm = 292 μm when CMP is the other substrate) as a phosphate donor. However, both UMP (Km = 153 μm) and CMP (Km = 266 μm) were equally acceptable as the phosphate acceptor. The optimal pH for the enzyme is 6.5. P1, P5-di(adenosine-5′) pentaphosphate was found to be a competitive inhibitor of both ATP and UMP. |
| تدمد: | 1532-2548 0032-0889 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::e1d9e8420c6fe454f77e91ad8898abc4 https://doi.org/10.1104/pp.117.1.245 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi...........e1d9e8420c6fe454f77e91ad8898abc4 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15322548 00320889 |
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