TAILIEUCHUNG - Báo cáo khoa học: Molecular characterization of recombinant mouse adenosine kinase and evaluation as a target for protein phosphorylation

The regulation of adenosine kinase (AK) activity has the potential to control intracellular and interstitial adenosine (Ado) concentrations. In an effort to study the role of AK in Ado homeostasis in the central nervous system, two iso-forms of the enzymewere cloned fromamouse brain cDNA library. Following overexpression in bacterial cells, the cor-responding proteins were purified to homogeneity. | Eur. J. Biochem. 271 3547-3555 2004 FEBS 2004 doi Molecular characterization of recombinant mouse adenosine kinase and evaluation as a target for protein phosphorylation Bogachan Sahin1 Janice W. Kansy1 Angus C. Nairn2 3 Jozef Spychala4 Steven E. Ealick5 Allen A. Fienberg3 6 Robert W. Greene1 7 and James A. Bibb1 1The University of Texas Southwestern Medical Center Dallas TX 2 Yale University School of Medicine New Haven CT The Rockefeller University New York NY 4University of North Carolina Chapel Hill NC 5Cornell University Ithaca NY 6Intra-Cellular Therapies Inc. New York NY 7Veterans Administration Medical Center Dallas TX USA The regulation of adenosine kinase AK activity has the potential to control intracellular and interstitial adenosine Ado concentrations. In an effort to study the role of AK in Ado homeostasis in the central nervous system two isoforms of the enzyme were cloned from a mouse brain cDNA library. Following overexpression in bacterial cells the corresponding proteins were purified to homogeneity. Both isoforms were enzymatically active and found to possess Km and Vmax values in agreement with kinetic parameters described for other forms of AK. The distribution of AK in discrete brain regions and various peripheral tissues was defined. To investigate the possibility that AK activity is regulated by protein phosphorylation a panel of protein kinases was screened for ability to phosphorylate recombinant mouse AK. Data from these in vitro phosphorylation studies suggest that AK is most likely not an efficient substrate for PKa pkg CaMKII CK1 CK2 MAPK Cdk1 or Cdk5. PKC was found to phosphorylate recombinant AK efficiently in vitro. Further analysis revealed however that this PKC-dependent phosphorylation occurred at one or more serine residues associated with the N-terminal affinity tag used for protein purification. Keywords adenosine kinase adenosine regulation protein serine threonine kinases CNS. Adenosine Ado

• Báo cáo khoa học
• Report medicine
• traditional medicine
• scientific reports
• intermediate cells
• cell proliferation
• medical analysis
• medical knowledge
• medical research
• analysis of cytoplasmic
• oxidation
• Crystal structure
• bacteria
• hemoglobin
• medicine
• breast cancer
• chemical reaction
• gastric cancer
• hormone medicine
• tumor cells
• biochemical studies
• environmental medicine