Cloning, characterization, and modeling of mouse and human guanylate kinases.
Guanylate kinase catalyzes the phosphorylation of either GMP to GDP or dGMP to dGDP and is an essential enzyme in nucleotide metabolism pathways. Despite its involvement in antiviral drug activation in humans and in mouse model systems and as a target for chemotherapy, the human and mouse primary structures have never been elucidated. Full-length cDNA clones encoding enzymatically active guanylate kinase were isolated from mouse B-cell lymphoma and human peripheral blood lymphocyte cDNA libraries. Multiple tissue Northern blots demonstrated an mRNA species of approximately 1 kilobase for both mice and humans in all tissue types examined. The mouse cDNA is predicted to encode a 198-amino acid protein with a molecular mass of 21,904 daltons. The human cDNA is predicted to encode a 197-amino acid protein with a molecular mass of 21,696 daltons. These proteins share 88% sequence identity with each other and 52-54% identity with the yeast guanylate kinase. Molecular modeling using the yeast diffraction coordinates indicates a high degree of conservation within the active site and maintenance of the overall structural integrity, despite a lack of similarity along the periphery of the enzyme.
Brady
W A
WA
Darwin Molecular Corporation, Bothell, Washington 98021, USA.
Kokoris
M S
MS
Fitzgibbon
M
M
Black
M E
ME
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D051528:Guanylate Kinases; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009703:Nucleoside-Phosphate Kinase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases