Modulation of adenine nucleotide concentrations in human plasma by erythrocytes and endothelial cells.
The regulation of plasma concentrations of adenine nucleotides is unsettled. We tested the possibility of extracellular adenosine triphosphate (ATP) production from adenosine diphosphate (ADP) at physiological low concentrations by erythrocytes and endothelial cells. Filtered erythrocytes and human umbilical vein endothelial cells (HUVEC) were incubated for 15 to 120 s with ADP (10 microM), supplemented with 3H-ADP (2.85 nM) or 14C-ADP (54.6 nM). Enzymatic conversion of ADP to ATP was detected by recovery of the radioactive label in the ATP fraction. ATP was measured in the supernatant using high performance liquid chromatography (HPLC) separation, scintillation techniques, and luminometry. Using etheno (epsilon)-labeled ADP (10 microM), the extracellular localization of the conversion was further corroborated. Following ADP application in plasma, no radioactivity was detected in the ATP fraction. However, in erythrocyte suspensions, 12.9% and 9.7% of the label were recovered in the ATP fraction after application of 3H- and 14C-ADP, respectively. Between 15 and 120 s after 3H-ADP application, the 3H-ATP fraction was found to be stable at around 10%. For the range of ADP concentrations studied (10-40 microM), no saturation of ATP production was achieved. The extracellular localization of conversion was supported by the recovery of the epsilon -label in the epsilon -ATP fraction. In contrast, on HUVEC a conversion of epsilon -ADP to epsilon -ATP was not observed. In conclusion, on erythrocytes there is rapid enzymatic conversion of extracellular ADP to ATP which may play a significant role in adjusting adenine nucleotide concentrations in human plasma. In endothelial cells, extracellular conversion of ADP to ATP is of quantitatively minor importance, if it contributes at all.
D000227:Adenine Nucleotides / Q000097:blood*; D000244:Adenosine Diphosphate / Q000097:blood; D000255:Adenosine Triphosphate / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D004730:Endothelium, Vascular / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D006801:Humans; D008297:Male; D010949:Plasma / Q000502:physiology*; D012016:Reference Values
D000227:Adenine Nucleotides; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate