Alkaline inorganic pyrophosphatase activity of mammalian-cell alkaline phosphatase |
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Authors: | Rody P. Cox Paul Gilbert Jun. Martin J. Griffin |
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Affiliation: | Department of Medicine, New York University School of Medicine, New York, and Summer Research Institute, Will Rogers Hospital, Saranac Lake, New York, N.Y., U.S.A. |
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Abstract: | Alkaline phosphatase prepared from mammalian cell cultures was found to have alkaline inorganic pyrophosphatase activity. Both of these activities appear to be associated with a single protein, as demonstrated by: (1) concomitant purification of alkaline phosphatase and alkaline inorganic pyrophosphatase; (2) proportional precipitation of alkaline phosphatase and inorganic pyrophosphatase activities by titrating constant amounts of an enzyme preparation with increasing concentration of antibody; (3) immune electrophoresis, which showed that precipitin bands that have alkaline phosphatase activity also have pyrophosphatase activity; (4) inhibition of pyrophosphatase activity by cysteine, an inhibitor of alkaline phosphatase activity; (5) similar subcellular localization of the two enzyme activities as demonstrated by histochemical methods; (6) hormonal and substrate induction of alkaline phosphatase activity in mammalian cell cultures, which produced a nearly parallel rise in inorganic pyrophosphatase activity. |
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