Studies on adenine and adenosine metabolism by intact human erythrocytes using high performance liquid chromatography.
Adenine and adenosine metabolism has been studied in intact human erythrocytes in vitro using high performance liquid chromatography, isotopic labeling and electrophoresis. Their metabolism to nucleotides was controlled by phosphoribose diphosphate synthesis which was phosphate dependent. Adenosine formed hypoxanthine or IMP depending upon Pi concentration, but adenosine kinase and deaminase activities were not affected by P levels. Free [14C]adenine and [14C]hypoxanthine were found in cellular extracts. Rapid interconversions occurred to give a distribution for ATP : ADP : AMP of 10 : 1 : 0.1. Marked decomposition of ATP to ADP and AMP occurred during incubations in plasma and Earle's media in air on nitrogen, but ATP levels remained stable in phosphate buffers and in the presence of oxygen. At physiological Pi (1 mM) adenosine kinase activity grossly exceeded adenine phosphoribosyltransferase activity. The latter was approximately 7 fold that of hypoxanthine phosphoribosyltransferase activity. These differences decreased with increasing Pi levels. No significant increase in corresponding nucleotides was obtained by incubation with high levels (0.5 mM) of adenine, guanine or guanosine at physiological Ii, ATP increased by 10% independently of the substrate employed and significant amounts of IMP and GTP were formed adenosine and guanosine, respectively. The existence of a bound intracellular pool of ATP is suggested.
D000225:Adenine / Q000097:blood*; D000241:Adenosine / Q000097:blood*; D000244:Adenosine Diphosphate / Q000097:blood; D000255:Adenosine Triphosphate / Q000097:blood; D001786:Blood Glucose / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000378:metabolism*; D006019:Glycolysis; D006801:Humans; D007292:Inosine Nucleotides / Q000097:blood; D007700:Kinetics; D007773:Lactates / Q000097:blood; D011687:Purines / Q000097:blood
D001786:Blood Glucose; D007292:Inosine Nucleotides; D007773:Lactates; D011687:Purines; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000225:Adenine; D000241:Adenosine