Partial characterization of folate uptake in microvillous membrane vesicles isolated from human placenta

ABSTRACT

The purpose of the present study was to determine biochemical parameters of folate uptake, and the putative contribution of the membrane-anchored folate receptor in microvillous membrane vesicles obtained from the syncytiotrophoblast of human term placenta. Uptake of [3H]-pteroylglutamic acid (PGA) by microvillous membrane vesicles was pH dependent with a maximum at pH 6.0, and attained equilibrium at 60 min of incubation. Uptake was higher in the presence of an inward pH gradient (pHout = 6.0; pHin = 7.5) than in the absence of the gradient (pHout = pHin = 6.0). The effect of changes in medium osmolality showed that both binding to the vesicular membrane and internalization contributed to the measured [3H]-PGA uptake. Equilibrium uptake experiments using [3H]-PGA concentrations within the physiological range of folate in blood serum showed that saturation was achieved at 30 nM and revealed a single class of binding sites with a Kd of 1.8 nM for [3H]-PGA. Cleavage of the glycosyl-phosphatidylinositol moiety of the folate receptor, which anchors the receptor to the membrane, with phosphatidylinositolspecific phospholipase C resulted in a reduction of about 80 per cent in [3H]-PGA uptake. In conclusion, our results showed that the folate uptake in the maternally facing membrane of the human placenta presents a saturable component and is mediated by the folate receptor to ensure an adequate maternal-fetal folate transfer.