PMID: 3769922Oct 1, 1986Paper

Selective preparation and characterization of membranous and soluble forms of alkaline phosphatase from rat tissues. A comparison with the serum enzyme

European Journal of Biochemistry
A MikiY Ikehara

Abstract

We developed a method for selective preparation of two forms of alkaline phosphatase from rat tissues. The enzyme was extracted by n-butanol treatment at pH 5.5 and pH 8.5 as soluble and aggregated (membranous) forms, respectively. The soluble form prepared from liver was found to be identical with the serum enzyme. Complete solubilization of the membrane-bound enzyme without detergents had a great advantage in its purification. Rat hepatoma AH-130 cells enriched in alkaline phosphatase were first used for purification of the liver-type enzyme. The hepatoma enzyme, purified by chromatographies on concanavalin-A-Sepharose, Sephacryl S-300 and hydroxyapatite was used for production of antibodies specific for the liver-type isozyme. An immunoaffinity column, prepared with anti-(hepatoma-enzyme) IgG was utilized for the enzyme purification from other tissues including the membranous form. Analyses of amino acid composition of the purified enzymes revealed that all the liver-type enzymes from hepatoma, liver, kidney and serum had the same composition, whereas the intestinal type consisted of the composition distinctly different from that in the liver type. In addition, there was no significant difference in amino acid composition be...Continue Reading

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Citations

Oct 1, 1989·Acta diabetologica latina·C GazzarriniB De Bernard
May 7, 1999·Applied Biochemistry and Biotechnology·A A PantazakiD A Kyriakidis
Mar 1, 1997·Archives of Gerontology and Geriatrics·A SafadiA Z Reznick
Feb 24, 2001·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·K HoshiH Ozawa
May 15, 2002·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Katsuhiro OnoNobuo Kugai
Jan 1, 1992·Archives Internationales De Physiologie, De Biochimie Et De Biophysique·M Gilles-Baillien, E Croux
Oct 13, 2004·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Isao KiiAkira Kudo
Sep 17, 2003·International Journal of Experimental Pathology·Jola J KapojosWinston W Bakker
Jul 18, 2002·European Journal of Oral Sciences·Keita TanabeHidehiro Ozawa

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