Ammonia permeability of the peribacteroid membrane (PBM) from N(2)-fixing soybean nodules was measured (8x10(-5) m/s) using isolated PBM in a stopped-flow spectrofluorimeter. Ammonia (NH(3)) uptake into PBM vesicles was inhibited by up to 42% by HgCl(2) (EC(50)=2.9 microM, mercaptoethanol-reversible) and reduced by ATP pre-incubation. The activation energy of NH(3) uptake (52 kJ/mol) increased (118 kJ/mol) with HgCl(2). Water transport was also HgCl(2)-sensitive (EC(50)=52.6 microM), but increased by ATP pre-incubation. NH(3) and H(2)O may permeate via different pathways through Nodulin 26 or there is another protein on the PBM that is permeable to NH(3).