Isolation and characterization of a cDNA clone encoding rat nucleoside diphosphate kinase.
A cDNA clone for cytosolic nucleoside diphosphate (NDP) kinase was isolated from a cDNA library of rat skeletal muscle using synthetic oligonucleotides as probes. The clone constitutes a 621-base pair cDNA sequence including the 456-base pair coding region and 137-base pair 3'-untranslated one with polyadenylation site. The complete primary structure of NDP kinase was deduced from the coding sequence. An NH2-terminal amino acid sequence analysis suggested that the translated enzyme protein suffered proteolytic cleavage followed by modification at the alpha-NH2 group of the newly produced NH2-terminal amino acid residue. Taking this into account, it was tentatively concluded that the mature NDP kinase consists of 147 amino acid residues with a molecular weight of 16,724. Northern blot hybridization analysis showed that NDP kinase mRNA could be detected in total RNA fractions of brain, spleen, heart, lung, liver, kidney, testis as well as skeletal muscle, and that there was no difference in the size of mRNAs from these tissues. Tissue distribution of the mRNA nearly paralleled those of protein moiety and activity of the enzyme.
Kimura
N
N
Department of Biochemistry, Tokyo Metropolitan Institute of Gerontology, Japan.
Shimada
N
N
Nomura
K
K
Watanabe
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D015723:Gene Library; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology*; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics*; D015345:Oligonucleotide Probes; D010770:Phosphotransferases / Q000235:genetics*; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D015345:Oligonucleotide Probes; D004247:DNA; D010770:Phosphotransferases; D009701:Nucleoside-Diphosphate Kinase