The transport of pteridines in CCRF-CEM human lymphoblastic cells.
The transport routes used by CCRF-CEM human lymphoblastoid cells for the influx and efflux of unconjugated pteridines were analyzed using [3H]6-hydroxymethylpterin as a model compound. Influx proceeds by a mechanism that exhibits a Km of 66.7 microM and a Vmax of 0.077 nmol/min per mg cellular protein. The process is somewhat sensitive to metabolic inhibitors, particularly uncouplers of oxidative phosphorylation, and is significantly affected by the presence of other pteridines in the extracellular medium. The results suggest that pterins with either no 6-substituent (pterin) or those with methyl, hydroxyl, or formyl groups in this position, which exhibit Ki values between 25 and 77 microM, may share the same pathway for uptake. 6-Carboxypterin exhibits low affinity for the system (Ki greater than 500 microM), as do 7-substituted and 6,7-di-substituted derivatives and compounds with larger groups at the 6-position, such as neopterin and biopterin (Ki = 250-300 microM). Efflux of [3H]6-hydroxymethylpterin occurs rapidly and can proceed by at least two routes. The first, comprising approximately 50% of total efflux, is inhibited by extracellular pterins and exhibits similar properties to the uptake system in both its pattern of sensitivity to metabolic inhibitors and its specificity for pteridine structure. The route by which the remaining efflux occurs is relatively insensitive to metabolic inhibition. Adenine significantly inhibits 6-hydroxymethylpterin influx and efflux (Ki = 10.6 microM for uptake) but does not appear to share the same transport system. Similarly, methotrexate and folic acid exhibit little affinity for the unconjugated pteridine transport routes.
Webber
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Department of Basic and Clinical Research, Scripps Clinic and Research Foundation, La Jolla, CA 92037.
Nazarbaghi
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D001692:Biological Transport; D002250:Carbon Radioisotopes; D002460:Cell Line; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D006801:Humans; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000378:metabolism*; D011621:Pteridines / Q000032:analysis / Q000037:antagonists & inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D011622:Pterins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000493:pharmacokinetics; D014316:Tritium; D014407:Tumor Cells, Cultured
D002250:Carbon Radioisotopes; D011621:Pteridines; D011622:Pterins; D014316:Tritium; C005926:2-amino-4-hydroxy-6-formylpteridine; C013044:6-hydroxymethylpterin