1,25-Dihydroxyvitamin D3-glycoside: Identification of a calcinogenic principle of solanum malacoxylon |
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Authors: | Mark R Haussler Robert H Wasserman Toni A McCain Meinrad Peterlik Kristine M Bursac Mark R Hughes |
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Institution: | 1. Department of Biochemistry University of Arizona Tucson, Arizona 85724, USA;1. Department of Physical Biology Cornell University Ithaca, New York 14853, USA |
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Abstract: | The water soluble calcinogenic factor present in the plant is partially purified by selective extraction and chromatography on silicic acid and then hydrolyzed with a mixed preparation of glycosidases from the sea worm, . Hydrolysis produces a chloroform soluble factor with biologic characteristics of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3), the hormonal form of vitamin D. Purification of this factor is accomplished by chromatography on Sephadex LH-20, silicic acid, and Celite columns, yielding 3 μg of active material. During the isolation, bioactivity (as assessed by the ability of fractions to compete with labeled 1,25-(OH)2D3 for binding to a specific intestinal receptor protein) migrates exactly with authentic tritiated 1,25-(OH)2D3. The purified factor has an ultraviolet absorption spectrum identical to that of 1,25-(OH)2D3 and analysis via direct probe mass spectrometry yields a parent molecular ion of m/e 416 and a fragmentation pattern indistinguishable from synthetic 1,25-(OH)2D3 hormone. We therefore conclude that the vitamin D-like principle in is a sterol-glycoside which contains the 1,25-(OH)2D3 molecule as its active sterol component. |
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