First Author | Ropp PA | Year | 1996 |
Journal | Arch Biochem Biophys | Volume | 336 |
Issue | 1 | Pages | 105-12 |
PubMed ID | 8951040 | Mgi Jnum | J:36993 |
Mgi Id | MGI:84399 | Doi | 10.1006/abbi.1996.0537 |
Citation | Ropp PA, et al. (1996) Cloning and expression of a cDNA encoding uridine kinase from mouse brain. Arch Biochem Biophys 336(1):105-12 |
abstractText | Uridine kinase is the rate-limiting enzyme in the pyrimidine salvage pathway of all mammalian cells. A cDNA for uridine kinase from mouse brain has been isolated, sequenced, and characterized. This is the first report of a complete nucleotide sequence for mammalian uridine kinase. The isolated cDNA is only 95% complete, missing the first 17 codons. The correct 5'-terminus sequence was obtained from high-stringency screening of a mouse liver genomic DNA library. The translated cDNA sequence encodes a protein of 277 amino acids (Mr 31,068). A truncated form of the cDNA was expressed in Escherichia coli. The expressed protein displayed uridine kinase activity and readily formed a tetramer, the most active form of the wild-type enzyme. Analysis of the amino acid sequence identified the three ATP-binding site consensus motifs. The predicted secondary structure for uridine kinase and the sequence comparison with three kinases having known crystal structures are consistent with uridine kinase having an alpha/beta core structure of the nucleotide-binding fold found in many kinases. We have also isolated and cloned a nonfunctional, processed pseudogene from mouse genomic DNA. This pseudogene sequence is 94% identical with the coding DNA. |