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• W2034846812 abstract "A study of the vitamin D3 (D3) 25-hydroxylase was undertaken in an in vivo-in vitro model. {3H}-D3 () was injected into the portal vein and the liver was excised 18 seconds later. The liver homogenate was then submitted to differential centrifugation and the amount of {3H}-D3 incorporated in the subcellular fractions was evaluated. The microsomal fraction was also incubated in vitro and the appearance of {3H}-25-hydroxyvitamin D3 [25(OH)D3] was determined by high performance liquid chromatography (HPLC). Results showed that the fractional liver {3H}-D3 uptake varied between 37 percent and 48 percent of the dose injected. The intracellular distribution of {3H}-D3 showed that most of the vitamin was incorporated into the microsomal fraction (45% to 50% of the intracellular {3H}-D3) except at the highest dose of {3H}-D3 where the cytosolic fraction contained the highest amount (56.4%) of the incorporated vitamin. Mathematical analysis of the intracellular {3H}-D3 distribution showed that the microsomal fraction was the only subcellular fraction that was found to incorporate {3H}-D3 in relation to the total liver uptake of the vitamin. The apparent Michaelis-Menten kinetics of the {3H}-D3-25-hydroxylase showed that with substrate concentration of up to 88.5 nM, the apparent Km and Vmax were 28.2 nM and 25.8 fentomoles (fmol) · min−1 · mg microsomal pro−1, respectively, but the reaction lost considerable efficiency with higher substrate concentrations. With the in vivo-in vitro model used, the cytosolic fraction was not essential for the optimal C-25 hydroxylation of D3. These results show that the endoplasmic reticulum of rat hepatocytes possess a high capacity for D3 incorporation. Moreover, the observed decreases in total liver and microsomal D3 sequestration as well as in enzyme efficiency at high D3 concentration suggest that all intrahepatic events may be interrelated in the reported constraints related to the liver production of 25(OH)D3." @default.
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• W2034846812 date "1985-03-01" @default.
• W2034846812 modified "2023-10-02" @default.
• W2034846812 title "Sequestration and microsomal C-25 hydroxylation of {3H}-vitamin D3 by the rat liver" @default.
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• W2034846812 doi "https://doi.org/10.1016/0026-0495(85)90007-1" @default.
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