Furosemide: a Specific Inhibitor of Pi Transport across the Plasma Membrane of Plant Cells

A search was made for inhibitors of P_i uptake that act directlyon the P_i transporter in the plasma membranes of Catharanthusroseus cells to inhibit P_i uptake without inhibition of protonpumping. Using standard electrodes, we monitored changes inpH and in the concentration of K⁺ ions, as well as the rateof P_i uptake, when an inhibitor to be tested was applied tothe cells in unbuffered medium. A9C (28 µM), a blockerof anion channels, inhibited P_i uptake but it also inhibitedthe proton pump. However, a structurally similar inhibitor,furosemide, inhibited P_i uptake without inhibiting proton pumping. It is suggested that the carboxylic group of these inhibitorsinteracts with the P_i-binding site (probably an amino group)of the P_i transporter in the plasma membrane and that the hydrophobicstructure of these inhibitors facilitates their accumulationin the plasma membrane. ³Present address: Department of Biology, Hitotsubashi University,2-1 Naka, Kunitachi, Tokyo, 186 Japan