The metabolism of phosphatidylinositol in the thyroid gland of the pig.
1. The metabolism of phosphatidylinositol in pig thyroid has been investigated as a basis for understanding the specific stimulation of the synthesis of this phospholipid in the gland by thyrotropin. 2. The gland contained an active Ca(2+)-dependent phosphatidylinositol-splitting enzyme with an optimum pH of 5.3-5.5. 3. The major water-soluble product (65%) formed by this catabolic enzyme was not phosphorylinositol but a related compound, which may be a cyclic phosphorylinositol. Both this and phosphorylinositol (35%) were released simultaneously from the phosphatidylinositol substrate. 4. The phosphatidylinositol-splitting enzyme was found almost exclusively in the supernatant fraction obtained by homogenization of the gland. It was not present in the acid-phosphatase-containing particulate fraction. 5. The incorporation of [2-(3)H(1)]inositol into phosphatidylinositol in the presence of either CDP-diglyceride or CTP+ATP was most active in the microsomal fraction. 6. When thyroidal microsomes were labelled with [(3)H]inositol and (32)P, and then incubated with unlabelled inositol, there was a dramatic loss of (3)H labelling from the phosphatidylinositol, which was not accompanied by an equivalent loss of (32)P from the phosphate moiety. This turnover of the inositol moiety required nucleotide coenzymes. It is postulated that the phosphatidylinositol is split into inositol and a phosphorus-containing lipid precursor of the phospholipid that remains on the microsomal membrane and is recycled. 7. Isolated thyroidal mitochondria synthesized phosphatidylinositol from [2-(3)H(1)]inositol only because of their contaminating microsomal component. 8. Some evidence has been obtained of a rapid transfer of phosphatidylinositol molecules from thyroidal microsomes to mitochondria when these were incubated together in the presence of a supernatant fraction. 9. Both phosphatidylinositol breakdown by the supernatant fraction of the gland and synthesis by the microsomes were totally inhibited by 1mm-chlorpromazine. This drug is known to suppress thyrotrophin-induced stimulation of activity in thyroid slices.
D000135:Acid Phosphatase / Q000032:analysis; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002458:Cell Fractionation; D002498:Centrifugation; D002746:Chlorpromazine / Q000494:pharmacology; D003597:Cytosine Nucleotides / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000032:analysis; D066298:In Vitro Techniques; D007294:Inositol / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D009708:Nucleotidases / Q000032:analysis; D010088:Oxidoreductases / Q000032:analysis; D010716:Phosphatidylinositols / Q000378:metabolism*; D010759:Phosphorus Isotopes; D013552:Swine; D013961:Thyroid Gland / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D014316:Tritium
D003597:Cytosine Nucleotides; D010716:Phosphatidylinositols; D010759:Phosphorus Isotopes; D014316:Tritium; D007294:Inositol; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D003576:Electron Transport Complex IV; D009708:Nucleotidases; D000135:Acid Phosphatase; D002746:Chlorpromazine