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  • Title: Transport and oxidation of choline by liver mitochondria.
    Author: Tyler DD.
    Journal: Biochem J; 1977 Sep 15; 166(3):571-81. PubMed ID: 23103.
    Abstract:
    1. Rapid choline oxidation and the onset of P(i)-induced swelling by liver mitochondria, incubated in a sucrose medium at or above pH7.0, required the addition of both P(i) and an uncoupling agent. Below pH7.0, P(i) alone was required for rapid choline oxidation and swelling. 2. Choline oxidation was inhibited by each of several reagents that also inhibited P(i)-induced swelling under similar conditions of incubation, including EGTA, mersalyl, Mg(2+), the Ca(2+)-ionophore A23187, rotenone and nupercaine. None of these reagents had any significant effect on the rate of choline oxidation by sonicated mitochondria. There was therefore a close correlation between the conditions required for rapid choline oxidation and for P(i)-induced swelling to occur, suggesting that in the absence of mitochondrial swelling the rate of choline oxidation is regulated by the rate of choline transport across the mitochondrial membrane. 3. Respiratory-chain inhibitors, uncoupling agents (at pH6.5) and ionophore A23187 caused a loss of endogenous Ca(2+) from mitochondria, whereas nupercaine and Mg(2+) had no significant effect on the Ca(2+) content. Inhibition of choline oxidation and mitochondrial swelling by ionophore A23187 was reversed by adding Ca(2+), but not by Mg(2+). It is concluded that added P(i) promotes the Ca(2+)-dependent activation of mitochondrial membrane phospholipase activity in respiring mitochondria, causing an increase in the permeability of the mitochondrial inner membrane to choline and therefore enabling rapid choline oxidation to occur. Nupercaine and Mg(2+) appear to block choline oxidation and swelling by inhibiting phospholipase activity. 4. Choline was oxidized slowly by tightly coupled mitochondria largely depleted of their endogenous adenine nucleotides, suggesting that these compounds are not directly concerned in the regulation of choline oxidation. 5. The results are discussed in relation to the possible mechanism of choline transport across the mitochondrial membrane in vivo and the influence of this process on the pathways of choline metabolism in the liver.
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