Mm. Gutierrez et Km. Giacomini, EXPRESSION OF A HUMAN RENAL SODIUM NUCLEOSIDE COTRANSPORTER IN XENOPUS-LAEVIS OOCYTES, Biochemical pharmacology, 48(12), 1994, pp. 2251-2253
In this study, Xenopus laevis oocytes injected with poly(A)(+) RNA (mR
NA) isolated from human kidney were used to express a Na+-nucleoside c
otransporter. Na+-stimulated [H-3]thymidine uptake was enhanced 2-3-fo
ld in oocytes injected with 50 ng poly(A)(+) RNA and 4-5-fold in oocyt
es injected with 20 ng of a size-fractionated human renal cortex mRNA
fragment (2-3 kb) in comparison with water-injected oocytes. Na+-depen
dent thymidine uptake in oocytes injected with the 2-3 kb mRNA fragmen
t was inhibited significantly by thymidine and guanosine but not by fo
rmycin B, consistent with the N4 Na+-nucleoside cotransporter. The K-m
(28 mu M) of Na+-dependent thymidine uptake in the oocytes injected w
ith the 2-3 kb mRNA fragment was similar to the K-m (27 mu M) of Na+-d
ependent thymidine uptake obtained in human renal brush border membran
e vesicles. These data suggest for the first time that a Na+-nucleosid
e cotransporter from human kidney can be expressed in X. laevis oocyte
s.