NUCLEOSIDE TRANSPORT IN CULTURED LLC-PK1 EPITHELIA

被引:18
作者
GRIFFITH, DA [1 ]
DOHERTY, AJ [1 ]
JARVIS, SM [1 ]
机构
[1] UNIV KENT,BIOL LAB,CANTERBURY CT2 7NJ,KENT,ENGLAND
基金
英国医学研究理事会;
关键词
NUCLEOSIDE TRANSPORT; FACILITATED TRANSPORT; ACTIVE TRANSPORT; NITROBENZYLTHIOINOSINE; LLC-PK1; EPITHELIUM; (PIG KIDNEY BRUSH-BORDER VESICLE);
D O I
10.1016/0005-2736(92)90010-J
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The transport of nucleosides by LLC-PK1 cells, a continuous epithelial cell line derived from pig kidney, was characterised. Uridine influx was saturable (apparent K(m) approximately 34-mu-M at 22-degrees-C) and inhibited by > 95% by nitrobenzylthioinosine (NBMPR), dilazep and a variety of purine and pyrimidine nucleosides. In contrast to other cultured animal cells, the NBMPR-sensitive nucleoside transporter in LLC-PK1 cells exhibited both a high affinity for cytidine (apparent K(i) approximately 65-mu-M for influx) and differential 'mobility' of the carrier (the kinetic parameters of equilibrium exchange of formycin B are greater than those for formycin B influx). An additional minor component of sodium-dependent uridine influx in LLC-PK, cells became detectable when the NBMPR-sensitive nucleoside transporter was blocked by the presence of 10-mu-M NBMPR. This active transport system was inhibited by adenosine, inosine and guanosine but thymidine and cytidine were without effect, inhibition properties identical to the N1 sodium-dependent nucleoside carrier in bovine renal outer cortical brush-border membrane vesicles (Williams and Jarvis (1991) Biochem. J. 274, 27-33). Late proximal tubule brush-border membrane vesicles of porcine kidney were shown to have a much reduced Na+-dependent uridine uptake activity compared to early proximal tubule porcine brush-border membrane vesicles. These results, together with the recent suggestion of thc late proximal tubular origin of LLC-PK1 cells, suggest that in vivo nucleoside transport across thc late proximal tubule cell may proceed mainly via a facilitated-diffusion process.
引用
收藏
页码:303 / 310
页数:8
相关论文
共 36 条
[1]   NUCLEOSIDE TRANSPORT IN WALKER-256 RAT CARCINOSARCOMA AND S49 MOUSE LYMPHOMA-CELLS - DIFFERENCES IN SENSITIVITY TO NITROBENZYLTHIOINOSINE AND THIOL REAGENTS [J].
BELT, JA ;
NOEL, LD .
BIOCHEMICAL JOURNAL, 1985, 232 (03) :681-688
[2]   ABSENCE OF BINDING-SITES FOR THE TRANSPORT INHIBITOR NITROBENZYLTHIOINOSINE ON NUCLEOSIDE TRANSPORT-DEFICIENT MOUSE LYMPHOMA-CELLS [J].
CASS, CE ;
KOLASSA, N ;
UEHARA, Y ;
DAHLIGHARLEY, E ;
HARLEY, ER ;
PATERSON, ARP .
BIOCHIMICA ET BIOPHYSICA ACTA, 1981, 649 (03) :769-777
[3]   NA+-DEPENDENT NUCLEOSIDE UPTAKE IN AN ESTABLISHED RENAL EPITHELIAL-CELL LINE, OK [J].
DOHERTY, AJ ;
JARVIS, SM .
BIOCHEMICAL SOCIETY TRANSACTIONS, 1990, 18 (06) :1246-1247
[4]  
GSTRAUNTHALER GJA, 1988, RENAL PHYSIOL BIOCH, V11, P1
[5]  
HULL RN, 1976, IN VITRO CELL DEV B, V12, P670, DOI 10.1007/BF02797469
[6]   NUCLEOSIDE TRANSPORT IN RAT ERYTHROCYTES - 2 COMPONENTS WITH DIFFERENCES IN SENSITIVITY TO INHIBITION BY NITROBENZYLTHIOINOSINE AND PARA-CHLOROMERCURIPHENYL SULFONATE [J].
JARVIS, SM ;
YOUNG, JD .
JOURNAL OF MEMBRANE BIOLOGY, 1986, 93 (01) :1-10
[7]   NUCLEOSIDE TRANSPORT IN HUMAN-ERYTHROCYTES - A SIMPLE CARRIER WITH DIRECTIONAL SYMMETRY IN FRESH CELLS, BUT WITH DIRECTIONAL ASYMMETRY IN CELLS FROM OUTDATED BLOOD [J].
JARVIS, SM ;
HAMMOND, JR ;
PATERSON, ARP ;
CLANACHAN, AS .
BIOCHEMICAL JOURNAL, 1983, 210 (02) :457-461
[8]   ERYTHROCYTE NUCLEOSIDE TRANSPORT - ASYMMETRICAL BINDING OF NITROBENZYLTHIOINOSINE TO NUCLEOSIDE PERMEATION SITES [J].
JARVIS, SM ;
MCBRIDE, D ;
YOUNG, JD .
JOURNAL OF PHYSIOLOGY-LONDON, 1982, 324 (MAR) :31-46
[9]   EFFECTS OF TEMPERATURE ON THE TRANSPORT OF NUCLEOSIDES IN GUINEA-PIG ERYTHROCYTES [J].
JARVIS, SM ;
MARTIN, BW .
CANADIAN JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, 1986, 64 (02) :193-198
[10]   EXPRESSION OF THE RABBIT INTESTINAL N2 NA+ NUCLEOSIDE TRANSPORTER IN XENOPUS-LAEVIS OOCYTES [J].
JARVIS, SM ;
GRIFFITH, DA .
BIOCHEMICAL JOURNAL, 1991, 278 :605-607