Cation coupling properties of human concentrative nucleoside transporters hCNT1, hCNT2 and hCNT3.
The SLC28 family of concentrative nucleoside transporter (CNT) proteins in mammalian cells contains members of two distinct phylogenic subfamilies. In humans, hCNT1 and hCNT2 belong to one subfamily, and hCNT3 to the other. All three CNTs mediate inwardly-directed Na(+)/nucleoside cotransport, and are either pyrimidine nucleoside-selective (hCNT1), purine nucleoside-selective (hCNT2), or broadly selective for both pyrimidine and purine nucleosides (hCNT3). While previous studies have characterized cation interactions with both hCNT1 and hCNT3, little is known about the corresponding properties of hCNT2. In the present study, heterologous expression in Xenopus oocytes in combination with radioisotope flux and electrophysiological techniques has allowed us to undertake a side-by-side comparison of hCNT2 with other hCNT family members. Apparent K (50) values for Na(+) activation were voltage-dependent, and similar in magnitude for all three transporters. Only hCNT3 was also able to couple transport of uridine to uptake of H(+). The Na(+)/nucleoside stoichiometry of hCNT2, as determined from both Hill coefficients and direct charge/flux measurements, was 1:1. This result was the same as for hCNT1, but different from that of hCNT3 (2:1). The charge-to-(22)Na(+) uptake stoichiometry was 1:1 for all three hCNTs. In parallel with their division into two separate CNT subfamilies, hCNT2 shares common cation specificity and coupling characteristics with hCNT1, which differ markedly from those of hCNT3.
Smith
Kyla M
KM
The Membrane Protein Research Group, Department of Physiology, University of Alberta, Alberta, Canada.
Slugoski
Melissa D
MD
Cass
Carol E
CE
Baldwin
Stephen A
SA
Karpinski
Edward
E
Young
James D
JD
D000818:Animals; D001692:Biological Transport; D006801:Humans; D006859:Hydrogen / Q000378:metabolism*; D007700:Kinetics; D026901:Membrane Transport Proteins / Q000378:metabolism*; D009705:Nucleosides / Q000378:metabolism; D009865:Oocytes; D011994:Recombinant Proteins / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D014981:Xenopus
D026901:Membrane Transport Proteins; D009705:Nucleosides; D011994:Recombinant Proteins; C065171:cif nucleoside transporter; D006859:Hydrogen; D012964:Sodium