Nucleotidase activities in soluble and membrane fractions of three different mammalian cell lines.
Soluble cytoplasmic and membrane fractions were prepared from three cultured mammalian cell lines: 3T3 mouse fibroblasts, V79 hamster lung cells, and human "Cherry" B-lymphoblastoid cells. By using relatively specific nucleotidase assays, together with a phosphotransferase assay, the activities of three different enzymes (low-Km nucleotidase, high-Km nucleotidase, and 5'(3')-nucleotidase) capable of dephosphorylating deoxyribonucleoside 5'-monophosphates were determined in these fractions. The three nucleotidases exist simultaneously in all cell lines, but their relative amounts showed large variations. The 5'(3')-nucleotidase dominated Cherry and 3T3 cells, while in V79 cells equal amounts of this enzyme and the high-Km nucleotidase were recovered. In the membrane fractions, the low-Km nucleotidase was the predominant enzyme. We found no evidence for cell-cycle control of any nucleotidase. We postulated earlier that substrate cycles, involving 5'-nucleotidases and deoxyribonucleoside kinases, provide a mechanism for the regulation of deoxyribonucleotide pools. We suggest that both the low-Km nucleotidase and the 5'(3)-nucleotidase are candidate enzymes for such cycles.
Höglund
L
L
Department of Biochemistry I, Medical Nobel Institute, Karolinska Institutet, Stockholm, Sweden.
Reichard
P
P
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D002453:Cell Cycle; D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology* / Q000648:ultrastructure; D002478:Cells, Cultured; D006224:Cricetinae; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D006801:Humans; D008168:Lung / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D051379:Mice; D009708:Nucleotidases / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism
D010770:Phosphotransferases; D009708:Nucleotidases