Summary
This paper describes a purification procedure and some properties of a nonspecific nucleoside phosphotransferase of chick embryo, an activity which catalyzes the transfer of the phosphate ester from a deoxyribonucleotide or a pyrimidine ribonucleotide to a deoxyribonucleoside acceptor.
The enzyme is very unstable to heat, dilution and dialysis and it is almost entirely inactivated by DEAE-cellulose chromatography or gel filtration. A marked enhancement in its stability is caused by numerous nucleotides. In these experiments at least 920-fold purification was obtained by using dTTP (50μm) as nucleotide protector.
The enzyme, purified in presence of dTTP, has a molecular weight about 270 000, an isoelectric point of 6.27, a pH optimum of 8.8 and is stable at 37 °C at least for 10 min.
In absence of nucleotide protector, nucleoside phosphotransferase is converted at 37 °C or by gel filtration in a very small active form with a lower molecular weight (about 30 000) and a pH optimum of 7.6.
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Bianchi, P. A., Crathorn, A. R. and Shooter, K. V., 1962. Biochim. Biophys. Acta 61, 728–735.
Bresnick, E., Mainigi, K. D., Buccino, R. and Burleson, S. S., 1970. Cancer Res. 30, 2502–2506.
Bresnick, E., Mainigi, K. D., Mayfield, E. D. and Morris, H. P., 1969. Cancer Res. 29, 1932–1936.
Maley, G. F., Lorenson, M. G. and Maley, F., 1965. Biochem. Biophys. Res. Commun. 18, 364–370.
Taylor, A. T., Stafford, M. A. and Jones, O. W., 1972. J. Biol. Chem. 247, 1930–1935.
Sneider, T. W., Potter, V. R. and Morris, H. P., 1969. Cancer Res. 29, 40–54.
Hatanaka, M., Twiddy, E. and Gilden, R., 1969. J. Virol. 4, 801–803.
Olsen, I. and Harris, G., 1975. Biochem. J. 146, 489–496.
Epstein, S., Esanu, C. and Ruben, M. S., 1969. Biochim. Biophys. Acta 186, 280–285.
Greengard, O. and Machovich, R., 1972. Biochim. Biophys. Acta 286, 382–388.
Masui, H. and Garren, L. D., 1971. J. Biol. Chem. 246, 5407–5413.
Georgatsos, J. C., 1967. Arch. Biochem. Biophys. 121, 619–624.
Tunis, M. and Chargaff, E., 1960. Biochim. Biophys. Acta 37, 257–266.
Tunis, M. and Chargaff, E., 1960. Biochim. Biophys. Acta 37, 267–273.
Shiosaka, T., Okuda, H. and Fujii, S., 1971. Biochim. Biophys. Acta 246, 171–183.
Deng, O. and Ives, D. H., 1972. Biochim. Biophys. Acta 277, 235–244.
Arima, T., Masaka, M., Shiosaka, T., Okuda, H. and Fujii, S., 1971. Biochim. Biophys. Acta 246, 184–193.
Tesoriere, G., Vento, R., Calvaruso, G. and Taibi, G., 1978. Experientia 34, 1268–1269.
Maley, G. F. and Maley, F., 1963. Arch. Biochem. Biophys. 101, 342–349.
Hurlbert, R. B., Schmitz, H., Brumm, A. F. and Potter, V. R., 1954. J. Biol. Chem. 209, 23–39.
Gillette, P. C. and Claycomb, W. C., 1974. Biochem. J. 142, 685–690.
Martin, R. G. and Ames, B. N., 1961. J. Biol. Chem. 236, 1372–1379.
Karlsson, C., Davies, H., Ohman, J. and Andersson, V., 1973. In “Analytical Thin-Layer Gel Electrofucusing in polyacrylamide gel”, pp. 1–13, LKB-Producter, Bromma, Sweden.
Vesterberg, O., 1972. Biochim. Biophys. Acta 257, 11–19.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., 1951. J. Biol. Chem. 193, 265–275.
Martin, J. B. and Doty, D. M., 1956. In “Methods of Biochemical Analysis” (David Glick, ed), Vol. 3, pp. 7–9, Interscience Publishers, New York.
Brunngraber, E. F. and Chargaff, E., 1967. J. Biol. Chem. 242, 4834–4840.
Brunngraber, E. F. and Chargaff, E., 1970. J. Biol. Chem. 245, 4825–4831.
Rodgers, R. and Chargaff, E., 1972. J. Biol. Chem. 247, 5448–5455.
Breitman, T. R., 1963. Biochim. Biophys. Acta 67, 153–155.
Maley, G. F. and Maley, F., 1968. J. Biol. Chem. 243, 4506–4512.
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Tesoriere, G., Vento, R., Calvaruso, G. et al. Nucleoside phosphotransferase of chick embryo. Mol Cell Biochem 25, 171–178 (1979). https://doi.org/10.1007/BF00235365
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DOI: https://doi.org/10.1007/BF00235365