Cell surface adenylate kinase activity regulates the F(1)-ATPase/P2Y (13)-mediated HDL endocytosis pathway on human hepatocytes.
We have previously demonstrated on human hepatocytes that apolipoprotein A-I binding to an ecto-F(1)-ATPase stimulates the production of extracellular ADP that activates a P2Y(13)-mediated high-density lipoprotein (HDL) endocytosis pathway. Therefore, we investigated the mechanisms controlling the extracellular ATP/ADP level in hepatic cell lines and primary cultures to determine their impact on HDL endocytosis. Here we show that addition of ADP to the cell culture medium induced extracellular ATP production that was due to adenylate kinase [see text] and nucleoside diphosphokinase [see text] activities, but not to ATP synthase activity. We further observed that in vitro modulation of both ecto-NDPK and AK activities could regulate the ADP-dependent HDL endocytosis. But interestingly, only AK appeared to naturally participate in the pathway by consuming the ADP generated by the ecto-F(1)-ATPase. Thus controlling the extracellular ADP level is a potential target for reverse cholesterol transport regulation.
Fabre
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INSERM U563, Département LML Bat C, Hôpital Purpan, BP 3048, 31024, Toulouse Cedex 03, France.
Vantourout
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P
Champagne
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E
Tercé
F
F
Rolland
C
C
Perret
B
B
Collet
X
X
Barbaras
R
R
Martinez
L O
LO
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000263:Adenylate Kinase / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D004705:Endocytosis; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism*; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000378:metabolism*
D003470:Culture Media; D008075:Lipoproteins, HDL; D018048:Receptors, Purinergic P2; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000263:Adenylate Kinase; D009701:Nucleoside-Diphosphate Kinase; D000251:Adenosine Triphosphatases