Interaction of N1,N12-diacetylspermine with polyamine transport systems of polarized porcine renal cell line LLC-PK1.
LLC-PK(1) cells grown on porous membrane filters were employed as a model system to explore the renal transport of polyamines. The polarity of LLC-PK(1) monolayers was confirmed by the exclusive appearance of a Na(+)-dependent alpha-methylglucoside transport system on the apical surface. The uptake of free polyamines from the basolateral side of monolayers was consistent with the existence of a single class of transport system, while the existence of two kinetically distinct polyamine transport systems with higher and lower affinities on apical membranes was suggested. The results of competition studies indicated that each of these transporters was able to interact with putrescine, spermidine and spermine. LLC-PK(1) cells incorporated monoacetylspermine from the apical surface of monolayers at about half the rate of spermine uptake. Monoacetylspermine inhibited spermidine uptake, indicating that free polyamine transport systems also recognized the monoacetylated derivative. In contrast, N(1),N(12)-diacetylspermine did not inhibit spermidine uptake, nor was it incorporated into the cells, indicating the absence of transport systems that recognize N(1),N(12)-diacetylspermine on the apical membranes of LLC-PK(1) cells. These results may be relevant as to our previous observation that the content of diacetylpolyamines in urine is relatively constant, and may explain the excellence of N(1),N(12)-diacetylspermine as a tumor marker.
Miki
Toshiaki
T
Department of Molecular Physiology and Medical Research and Development Center, The Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, Tokyo 113-8613, Japan.
Hiramatsu
Kyoko
K
Kawakita
Masao
M
D000818:Animals; D001692:Biological Transport; D014408:Biomarkers, Tumor / Q000493:pharmacokinetics; D016764:Cell Polarity; D018374:LLC-PK1 Cells / Q000166:cytology / Q000378:metabolism*; D011073:Polyamines / Q000493:pharmacokinetics*; D013096:Spermine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D013552:Swine
D014408:Biomarkers, Tumor; D011073:Polyamines; D013096:Spermine; C029453:N',N''-diacetylspermine