PHOSPHORUS METABOLISM IN HUMAN ERYTHROCYTE:V. INCORPORATION OF P32 DURING VERY SHORT TIME INTERVALS

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As an attempt to elucidate the mechanism of phosphate uptake by human erythrocyte, P32 incorporation into acid-soluble organic phosphates during very short time period of incubation with added P32 orthophosphate was pursued by the use of ion exchange resin chromatography. The P32 incorporation into 2, 3-diphosphoglycerate was found to be a rather sluggish one. In an incubation time up to 90 seconds no detectable labeling of the ester was obtained. Even after 30 minutes of incubation the specific activity of 2, 3-diphosphoglyceric acid was several times as small as that of ATP. ATP and ADP became labeled to a detectable extent as early as 10 seconds after the addition of P32, when neither sugar phosphates nor phosphoglycerate took up practically no P32. The incorporation of P32 into ATP increases with time linearly with a very short lag. This suggests that there is an intermeniary step between inorganic orthophosphate in plasma and ATP in the cell. In addition to ATP, and ADP, several unidentified compounds with very early incorporation of P32 were found. However, their biological significance is ambiguous because they were present some times even in zero-time controls. A very rapid isotopic equilibration among the labile phosphate groups of ATP and ADP was demonstrated.
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