Uptake and metabolism of putrescine in confluent LLC-PK1 cells.
Putrescine is taken up by confluent pig kidney (LLC-PK1) cells at roughly equal rates over both Na(+)-dependent and Na(+)-independent pathways. The former is sensitive to 1 mM amiloride, but the latter is not. Uptake rates are similar at both the apical and basolateral surfaces. The principal fate of the putrescine is oxidative deamination, yielding a product that appears to be either gamma-aminobutyraldehyde or delta 1-pyrroline. Most of the remainder is converted to products tentatively identified as spermidine, spermine, or another unidentified product; these products as well as putrescine itself are lost from the cell at either surface. Changing the extracellular pH in the range of 6.8-8.0 has no affect on putrescine uptake. Cells acidified to intracellular pH 6.8 show a reduced capacity to incorporate radioactivity, an effect that may be due to inhibition of diamine oxidase. Depletion of ATP stores by treating cells with 2-deoxy-D-glucose and NaN3 does not reduce putrescine uptake, suggesting that the mechanism is not a primary active transporter. The Na(+)-dependent component of uptake is inhibited by 5-50 microM Hg2+ in a dose-dependent manner. p-Chloromercuribenzene sulfonic acid (p-CMBS) at high concentrations (500-1,000 microM) does not affect Na(+)-independent uptake but in the presence of Na+ depresses total uptake more than Na+ depletion alone, suggesting that Na+ enhances the binding of p-CMBS to both transporters. Spermidine and spermine compete with putrescine for uptake, but a variety of other organic bases and amino acids do not, indicating that polyamines are transported by mechanisms distinct from the transporters for those other compounds.
Hauser
M R
MR
University of Tennessee-Oak Ridge Graduate School of Biomedical Sciences.
Cook
J S
JS
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000584:Amiloride / Q000494:pharmacology; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D007668:Kidney; D007700:Kinetics; D011073:Polyamines / Q000494:pharmacology; D011700:Putrescine / Q000378:metabolism*; D013552:Swine
D000596:Amino Acids; D011073:Polyamines; D002731:4-Chloromercuribenzenesulfonate; D000584:Amiloride; D011700:Putrescine