Molecular cloning and functional expression of cDNAs encoding a human Na+-nucleoside cotransporter (hCNT1)

被引:184
作者
Ritzel, MWL
Yao, SYM
Huang, MY
Elliott, JF
Cass, CE
Young, JD
机构
[1] UNIV ALBERTA, DEPT PHYSIOL, MEMBRANE TRANSPORT GRP, EDMONTON, AB T6G 2H7, CANADA
[2] UNIV ALBERTA, DEPT MED MICROBIOL & IMMUNOL, EDMONTON, AB T6G 2H7, CANADA
[3] UNIV ALBERTA, DEPT BIOCHEM & ONCOL, EDMONTON, AB T6G 2H7, CANADA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY | 1997年 / 272卷 / 02期
关键词
kidney nucleoside transport system N2/cit; Xenopus oocyte; adenosine; deoxyadenosine; 3'-azido-3'-deoxythymidine;
D O I
10.1152/ajpcell.1997.272.2.C707
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We report identification of a new human nucleoside transporter protein by molecular cloning and functional expression of its cDNA. Previously, we used expression selection in Xenopus oocytes to isolate a cDNA from rat jejunal epithelium encoding the pyrimidine-selective Na+-dependent nucleoside transporter rCNT1 (Q.-Q. Huang, S. Y. M. Yao, M. W. L. Ritzel, A, R. P. Paterson, C. E. Cass, and J. D. Young. J. Biol. Chem. 269: 17757-17760, 1994). cDNAs for a human homologue of rCNT1, designated hCNT1, have been isolated from human kidney by hybridization cloning and reverse transcriptase polymerase chain reaction amplification strategies. hCNT1 was 83% identical to rCNT1 in amino acid sequence and exhibited the transport characteristics of an Na+-dependent nucleoside transporter with selectivity for pyrimidine nucleosides and adenosine when expressed in Xenopus oocytes. Deoxyadenosine, which undergoes net renal secretion, and guanosine were poor permeants. hCNT1 did, however, transport 3'-azido-3'-deoxythymidine. This is the first demonstration that members of the CNT family exist in human cells and provides evidence of their involvement in the renal transport of physiological nucleosides and nucleoside drugs. The hCNT1 gene was mapped to chromosome 15q25-26.
引用
收藏
页码:C707 / C714
页数:8
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