Biochim Biophys Acta 1999 Jan 12;14161-2:109-18.

Nucleobase transport in opossum kidney epithelial cells and Xenopus laevis oocytes: the characterisation, structure-activity relationship of uracil analogues and oocyte expression studies of sodium-dependent and -independent hypoxanthine uptake.

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The characteristics of hypoxanthine transport were examined in opossum kidney (OK) epithelial cells and Xenopus laevis oocytes. In both cell types hypoxanthine influx was mediated by two distinct transport systems: a high-affinity Na+-dependent system and a Na+-independent transporter. Na+-dependent hypoxanthine transport in OK cells was saturable (Km 0.78+/-0.29 microM) and was inhibited by guanine, uracil, thymine and 5-fluorouracil (Ki values 0.5-7 microM), whereas adenine had no effect. Substitutions at the 2- and 4-position had a marked effect on the ability of uracil to inhibit Na+/hypoxanthine influx by OK cells revealing that an oxo group at both the 2- and 4-positions of uracil is required for interacting with the transporter. The properties of Na+-dependent hypoxanthine influx in oocytes were similar to those observed in OK cells. In particular, xanthine and oxypurinol inhibited hypoxanthine influx, a characteristic not observed previously for the Na+/nucleobase carrier in pig LLC-PK1 renal cells. Na+-independent hypoxanthine influx in OK cells and oocytes was of a lower affinity (Km 90-180 microM). Adenine and guanine inhibited Na+-independent hypoxanthine flux in OK cells, but had no effect in oocytes. Injection of LLC-PK1 mRNA into oocytes resulted in a 1.5-fold stimulation of Na+/hypoxanthine flux over water-injected oocytes. These results reveal further heterogeneity in Na+/nucleobase cotransporters.

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Species referenced: Xenopus laevis
Genes referenced: pklr