Cloning and Expression of the Adenosine Kinase Gene from Rat and Human Tissues
Adenosine kinase is ubiquitous in eukaryotes and is a key enzyme in the regulation of the intracellular levels of adenosine, an important physiological effector of many cells and tissues. In this paper we report the cloning of cDNAs encoding adenosine kinase from both rat and human tissues. Two dist...
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Published in | Biochemical and biophysical research communications Vol. 231; no. 3; pp. 645 - 650 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
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United States
Elsevier Inc
24.02.1997
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Abstract | Adenosine kinase is ubiquitous in eukaryotes and is a key enzyme in the regulation of the intracellular levels of adenosine, an important physiological effector of many cells and tissues. In this paper we report the cloning of cDNAs encoding adenosine kinase from both rat and human tissues. Two distinct forms of adenosine kinase mRNA were identified in human tissues. Sequence variation between the two forms is restricted to the extreme 5′-end of the adenosine kinase mRNA, including a portion of the coding region, and is consistent with differential splicing of a single transcriptional product. We have expressed both forms inE. coliand produced soluble active enzyme which catalyzes the phosphorylation of adenosine with high specific activityin vitroand is susceptible to known adenosine kinase inhibitors. |
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AbstractList | Adenosine kinase is ubiquitous in eukaryotes and is a key enzyme in the regulation of the intracellular levels of adenosine, an important physiological effector of many cells and tissues. In this paper we report the cloning of cDNAs encoding adenosine kinase from both rat and human tissues. Two distinct forms of adenosine kinase mRNA were identified in human tissues. Sequence variation between the two forms is restricted to the extreme 5′-end of the adenosine kinase mRNA, including a portion of the coding region, and is consistent with differential splicing of a single transcriptional product. We have expressed both forms inE. coliand produced soluble active enzyme which catalyzes the phosphorylation of adenosine with high specific activityin vitroand is susceptible to known adenosine kinase inhibitors. Adenosine kinase is ubiquitous in eukaryotes and is a key enzyme in the regulation of the intracellular levels of adenosine, an important physiological effector of many cells and tissues. In this paper we report the cloning of cDNAs encoding adenosine kinase from both rat and human tissues. Two distinct forms of adenosine kinase mRNA were identified in human tissues. Sequence variation between the two forms is restricted to the extreme 5'-end of the adenosine kinase mRNA, including a portion of the coding region, and is consistent with differential splicing of a single transcriptional product. We have expressed both forms in E. coli and produced soluble active enzyme which catalyzes the phosphorylation of adenosine with high specific activity in vitro and is susceptible to known adenosine kinase inhibitors. |
Author | McNally, Teresa Meuth, Joseph L. Dorwin, Sarah A. Klute, Kimberly A. Cowart, Marlon Helfrich, Rosalind J. Simmer, Robert L. Halbert, Donald N. Idler, Kenneth B. Kowaluk, Elizabeth A. |
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BackLink | https://www.ncbi.nlm.nih.gov/pubmed/9070863$$D View this record in MEDLINE/PubMed |
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Cites_doi | 10.1073/pnas.93.3.1232 10.1016/0005-2744(80)90138-2 10.1111/j.1432-1033.1985.tb09110.x 10.1515/bchm2.1971.352.1.328 10.1146/annurev.bi.47.070178.003255 10.1021/bi00381a030 10.1016/S0021-9258(19)70780-7 10.1126/science.2448875 10.1016/S0021-9258(17)39669-2 10.1016/S0021-9258(17)30227-2 10.1016/S0021-9258(17)30226-0 10.1016/0003-2697(83)90418-9 10.1016/S0021-9258(18)45602-5 10.1093/nar/17.8.2919 10.1073/pnas.85.23.8998 10.1016/S0021-9258(19)86498-0 10.1016/0005-2744(81)90105-4 |
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SubjectTerms | Adenosine Kinase - genetics Adenosine Kinase - metabolism Amino Acid Sequence Animals Base Sequence Brain - enzymology Cloning, Molecular DNA, Complementary - genetics Humans Molecular Sequence Data Rats Recombinant Proteins - metabolism Structure-Activity Relationship |
Title | Cloning and Expression of the Adenosine Kinase Gene from Rat and Human Tissues |
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