Inhibitory effects of prostaglandin A1 on membrane transport of folates mediated by both the reduced folate carrier and ATP-driven exporters

Yehuda G. Assaraf, Esteban E. Sierra, Solomon Babani, I. David Goldman

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Studies are reported that describe the multifaceted inhibitory effects of prostaglandin A1 (PGA1) on processes that govern the transport of folates across the plasma membrane of Chinese hamster ovary (CHO) cells: the reduced folate carrier, RFC1, and ATP-dependent exporters. PGA1 was a noncompetitive inhibitor of MTX influx mediated by RFC1 with a K(i) of ~21 μM. The onset of inhibition was virtually instantaneous, not reversible, and appeared to require the incorporation of PGA1 into the lipid membrane; surface adsorption alone was insufficient for inhibition of RFC1 transport activity. In contrast, the effect of PGA1 on folic acid transport was small (~20% inhibition of total influx), consistent with the observation that the major portion of folic acid transport in CHO cells is mediated by a low pH mechanism distinct from RFC1. PGA1 was also a potent inhibitor of the ATP-driven efflux of both MTX and folic acid. At a concentration of 7 μM PGA1, the efflux rate constants for these folates were depressed by ~70 and ~50%, respectively. The net effects of PGA1 on the bidirectional folate fluxes translated into marked alterations in net transport. The addition of 7 μM PGA1 to cells at steady state with 1 μM MTX produced a rapid onset of net uptake and the achievement of an ~3-fold increase in the steady-state free MTX level as compared with untreated CHO cells. The addition of 7 μM PGA1 to cells at steady state with l μM folic acid produced an ~5-fold increase in the free folate level. These studies establish PGA1 as a potent inhibitor of both the reduced folate carrier and ATP-driven folate exporter(s). The noncompetitive nature of the inhibition of RFC1 is unique among anionic compounds, which are usually competitive inhibitors of the carrier. Copyright (C) 1999 Elsevier Science Inc.

Original languageEnglish (US)
Pages (from-to)1321-1327
Number of pages7
JournalBiochemical Pharmacology
Volume58
Issue number8
DOIs
StatePublished - Oct 15 1999

Fingerprint

Reduced Folate Carrier Protein
Folic Acid
Adenosine Triphosphate
Membranes
Cricetulus
Ovary
prostaglandin A1
Cell membranes
Membrane Lipids
Adsorption
Rate constants

Keywords

  • (anti)folate transport
  • Efflux transporters
  • Prostaglandin A
  • Reduced folate carrier

ASJC Scopus subject areas

  • Pharmacology

Cite this

Inhibitory effects of prostaglandin A1 on membrane transport of folates mediated by both the reduced folate carrier and ATP-driven exporters. / Assaraf, Yehuda G.; Sierra, Esteban E.; Babani, Solomon; Goldman, I. David.

In: Biochemical Pharmacology, Vol. 58, No. 8, 15.10.1999, p. 1321-1327.

Research output: Contribution to journalArticle

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abstract = "Studies are reported that describe the multifaceted inhibitory effects of prostaglandin A1 (PGA1) on processes that govern the transport of folates across the plasma membrane of Chinese hamster ovary (CHO) cells: the reduced folate carrier, RFC1, and ATP-dependent exporters. PGA1 was a noncompetitive inhibitor of MTX influx mediated by RFC1 with a K(i) of ~21 μM. The onset of inhibition was virtually instantaneous, not reversible, and appeared to require the incorporation of PGA1 into the lipid membrane; surface adsorption alone was insufficient for inhibition of RFC1 transport activity. In contrast, the effect of PGA1 on folic acid transport was small (~20{\%} inhibition of total influx), consistent with the observation that the major portion of folic acid transport in CHO cells is mediated by a low pH mechanism distinct from RFC1. PGA1 was also a potent inhibitor of the ATP-driven efflux of both MTX and folic acid. At a concentration of 7 μM PGA1, the efflux rate constants for these folates were depressed by ~70 and ~50{\%}, respectively. The net effects of PGA1 on the bidirectional folate fluxes translated into marked alterations in net transport. The addition of 7 μM PGA1 to cells at steady state with 1 μM MTX produced a rapid onset of net uptake and the achievement of an ~3-fold increase in the steady-state free MTX level as compared with untreated CHO cells. The addition of 7 μM PGA1 to cells at steady state with l μM folic acid produced an ~5-fold increase in the free folate level. These studies establish PGA1 as a potent inhibitor of both the reduced folate carrier and ATP-driven folate exporter(s). The noncompetitive nature of the inhibition of RFC1 is unique among anionic compounds, which are usually competitive inhibitors of the carrier. Copyright (C) 1999 Elsevier Science Inc.",
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N2 - Studies are reported that describe the multifaceted inhibitory effects of prostaglandin A1 (PGA1) on processes that govern the transport of folates across the plasma membrane of Chinese hamster ovary (CHO) cells: the reduced folate carrier, RFC1, and ATP-dependent exporters. PGA1 was a noncompetitive inhibitor of MTX influx mediated by RFC1 with a K(i) of ~21 μM. The onset of inhibition was virtually instantaneous, not reversible, and appeared to require the incorporation of PGA1 into the lipid membrane; surface adsorption alone was insufficient for inhibition of RFC1 transport activity. In contrast, the effect of PGA1 on folic acid transport was small (~20% inhibition of total influx), consistent with the observation that the major portion of folic acid transport in CHO cells is mediated by a low pH mechanism distinct from RFC1. PGA1 was also a potent inhibitor of the ATP-driven efflux of both MTX and folic acid. At a concentration of 7 μM PGA1, the efflux rate constants for these folates were depressed by ~70 and ~50%, respectively. The net effects of PGA1 on the bidirectional folate fluxes translated into marked alterations in net transport. The addition of 7 μM PGA1 to cells at steady state with 1 μM MTX produced a rapid onset of net uptake and the achievement of an ~3-fold increase in the steady-state free MTX level as compared with untreated CHO cells. The addition of 7 μM PGA1 to cells at steady state with l μM folic acid produced an ~5-fold increase in the free folate level. These studies establish PGA1 as a potent inhibitor of both the reduced folate carrier and ATP-driven folate exporter(s). The noncompetitive nature of the inhibition of RFC1 is unique among anionic compounds, which are usually competitive inhibitors of the carrier. Copyright (C) 1999 Elsevier Science Inc.

AB - Studies are reported that describe the multifaceted inhibitory effects of prostaglandin A1 (PGA1) on processes that govern the transport of folates across the plasma membrane of Chinese hamster ovary (CHO) cells: the reduced folate carrier, RFC1, and ATP-dependent exporters. PGA1 was a noncompetitive inhibitor of MTX influx mediated by RFC1 with a K(i) of ~21 μM. The onset of inhibition was virtually instantaneous, not reversible, and appeared to require the incorporation of PGA1 into the lipid membrane; surface adsorption alone was insufficient for inhibition of RFC1 transport activity. In contrast, the effect of PGA1 on folic acid transport was small (~20% inhibition of total influx), consistent with the observation that the major portion of folic acid transport in CHO cells is mediated by a low pH mechanism distinct from RFC1. PGA1 was also a potent inhibitor of the ATP-driven efflux of both MTX and folic acid. At a concentration of 7 μM PGA1, the efflux rate constants for these folates were depressed by ~70 and ~50%, respectively. The net effects of PGA1 on the bidirectional folate fluxes translated into marked alterations in net transport. The addition of 7 μM PGA1 to cells at steady state with 1 μM MTX produced a rapid onset of net uptake and the achievement of an ~3-fold increase in the steady-state free MTX level as compared with untreated CHO cells. The addition of 7 μM PGA1 to cells at steady state with l μM folic acid produced an ~5-fold increase in the free folate level. These studies establish PGA1 as a potent inhibitor of both the reduced folate carrier and ATP-driven folate exporter(s). The noncompetitive nature of the inhibition of RFC1 is unique among anionic compounds, which are usually competitive inhibitors of the carrier. Copyright (C) 1999 Elsevier Science Inc.

KW - (anti)folate transport

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