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Adenine phosphoribosyltransferase and hypoxanthine-guanine phosphoribosyltransferase immunoprecipitation reactions in human-mouse and human-hamster cell hybrids

Summary

Male New Zealand White rabbits were immunized with human adenine phosphoribosyltransferase (APRT) and hypoxanthine-guanine phosphoribosyltransferase (HGPRT), which were purified about 2000-fold and 800-fold, respectively, from erythrocytes by DEAE-cellulose chromatography, ammonium sulfate precipitation and preparative polyacrylamide gel electrophoresis. Specific immunoprecipitations of APRT and HGPRT were achieved with the antisera that were obtained and by using polyethylene glycol as a substitute for goat anti-(rabbit) gamma globulin. The activities of the human forms of these enzymes, whether from red blood cells or from cultured cells, were almost completely eliminated under the conditions of immunoprecipitation used. Little or no redution of APRT and HGPRT activities from mouse and Chinese hamster cells was observed. This discriminatory capacity of the antisera was successfully used for the identification of human APRT and HGPRT in human-mouse and human-hamster cell hybrids using the immunoprecipitation reaction.

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Abbreviations

APRT:

adenine phosphoribosyltransferase, E.C. 2.4.2.7

HGPRT:

hypoxanthine-guanine phosphoribosyltransferase, E.C. 2.4.2.8 (E.C. numbers from Enzyme Nomenclature: Recommendations of 1964, 1965)

PRPP:

5-phosphorylribose-1-pyrophosphate

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Held, K.R., Kahan, B. & DeMars, R. Adenine phosphoribosyltransferase and hypoxanthine-guanine phosphoribosyltransferase immunoprecipitation reactions in human-mouse and human-hamster cell hybrids. Humangenetik 30, 23–34 (1975). https://doi.org/10.1007/BF00273628

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Keywords

  • Ammonium Sulfate
  • Polyethylene Glycol
  • Zealand White Rabbit
  • Sulfate Precipitation
  • Ammonium Sulfate Precipitation