Multidrug resistance proteins (MRPs) are transporters for metabolic waste and xenobiotics and are known to export a wide range of substances from renal tubular cells. This study aimed to define and characterize the transport of mercuric conjugates of glutathione (GSH-Hg-GSH) in inside-out membrane vesicles containing MRP3 and MPR5. The functionality of the MRP3 and MRP5 vesicles was confirmed by measuring the uptake of [H]-estradiol and 5-6-carboxy-2',7'-dichloro-fluorescein (CDCF) over time (at 1, 5, 15, and 30 min). The uptake of GSH-Hg-GSH, containing radioactive mercury ([Hg]), was measured in each set of membrane vesicles over time, and the findings suggest that GSH-Hg-GSH is a substrate of MRP3 and MRP5. The saturation kinetics were also analyzed by measuring the uptake of 10 µM GSH-[Hg]-GSH in the presence of 25, 50, or 100 µM unlabeled GSH-Hg-GSH for 5 min at 37 °C. The transport of GSH-Hg-GSH by MRP3 ( = 25.6 µM; = 2.8 µM) and MRP5 ( = 32.9 µM; = 4.9 µM) was saturable. These findings are the first to show that MRP3 and MRP5 are capable of mediating the export of any form of mercury.