Evidence for the transport function of uricase, an oxidative enzyme.
[2-14C]urate uptake was examined in proteoliposomes prepared with phosphatidylcholine and either pig liver uricase or albumin, and in protein-free liposomes. Urate uptake was only evident in proteoliposomes that contained active uricase. Uptakes were indistinguishable in the presence and absence of inwardly directed gradients of sodium, potassium, or choline chloride or outwardly directed hydroxyl gradients. Both urate and allantoin accumulated within proteoliposomes during urate uptake; however, [2-14C]allantoin was not taken up by proteoliposomes. Urate uptake was accelerated in the presence of unlabeled urate in the trans position, saturable, and competitively inhibited by oxonate, findings consistent with carrier-mediated transport. Finally, the kinetics of urate uptake and oxidation were virtually identical, implying that the transporter is uricase. Thus, these studies provide evidence that uricase can function as a transport protein for urate when inserted in a lipid bilayer: transport via uricase is neither cation dependent (not a cotransporter) nor dependent on an exchangeable anion (not a urate/anion exchanger). Additionally, these studies demonstrate that neither urate nor allantoin cross lipid bilayers by simple or nonionic diffusion.
Pordy
W T
WT
Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029.
Lipkowitz
M S
MS
Abramson
R G
RG
D000481:Allantoin / Q000378:metabolism; D000818:Animals; D001693:Biological Transport, Active; D002794:Choline / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008854:Microscopy, Electron; D010713:Phosphatidylcholines / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D011510:Proteolipids; D012964:Sodium / Q000378:metabolism; D013552:Swine; D014503:Urate Oxidase / Q000378:metabolism*
D010713:Phosphatidylcholines; D011510:Proteolipids; C020485:proteoliposomes; D000481:Allantoin; D012964:Sodium; D014503:Urate Oxidase; D002794:Choline; D011188:Potassium