ADENOSINE KINASE OF MAMMALIAN BRAIN: PARTIAL PURIFICATION AND ITS ROLE FOR THE UPTAKE OF ADENOSINE

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Abstract
Abstract— Adenosine metabolism in the homogenate of brain mainly undergoes deamination to inosine and hypoxanthine, while uniformly labelled [14C]adenosine injected into the carotid artery or [8‐14C]adenosine incubated with brain slices was mostly phosphorylated to [14C]adenine nucleotides in brain cells. Adenosine kinase has now been partially purified from homogenates of guinea pig brain. The kinase preparation was free of adenosine deaminase, almost free of adenosine triphosphatase and had a Km of the order of 2 × 10‐5M for adenosine.Kinetic studies with brain slices showed that adenosine reached the cells by diffusion and that the diffusion was facilitated by subsequent phosphorylation to adenine nucleotides. From the following experimental results, it is concluded that the phosphorylation is catalysed by adenosine kinase quantitatively. (1) During the uptake and phosphorylation of adenosine by brain slices, the nucleoside did not split to adenine and ribose moieties. (2) The rate of formation of adenine nucleotides in the slices was a hyperbolic function of the concentration of adenosine in the medium, showing an apparent Km foradenosine of the order of 2 × 10‐5 M. (3) Some analogues of adenosine inhibited both the facilitated diffusion of adenosine and the kinase activity, but ouabain (0.005 mM) did not inhibit either.

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