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Purification and Characterization of Hypoxanthine-Guanine Phosphoribosyl Transferase from Cultured HTC Cells

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Purine Metabolism in Man—II

Abstract

Hypoxanthine-guanine phosphoribosyl transferase (HGPRTase.) from |35S|-methionyl-labelled rat hepatoma (HTC) cells has been purified more than 600-fold to near homogeneity by making use of conventional and affinity chromatographic techniques (Table 1). The conventional purification procedure utilizes a pH 5.0 precipitation, an ammonium sulfate precipitation, DEAE-cellulose chromatography, and heating at 80–85°according to the method of Olsen and Milman, 1974. Using this procedure, a partially pure enzyme preparation is obtained. Affinity chromatography according to Hughes et al., 1975, can be used either before or after the DEAE step to yield pure enzyme.

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References

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© 1977 Plenum Press, New York

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Zannis, V.I., Gudas, L.J., Doyle, D., Martin, D.W. (1977). Purification and Characterization of Hypoxanthine-Guanine Phosphoribosyl Transferase from Cultured HTC Cells. In: Müller, M.M., Kaiser, E., Seegmiller, J.E. (eds) Purine Metabolism in Man—II. Advances in Experimental Medicine and Biology. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-4223-6_21

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  • DOI: https://doi.org/10.1007/978-1-4613-4223-6_21

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-4225-0

  • Online ISBN: 978-1-4613-4223-6

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