Transport of organic cations by kidney epithelial cell line LLC-PK1

Transport of two organic cations, N1-methylnicotinamide (NMN) and tetraethylammonium (TEA), was investigated in LLC-PK1 cells grown on Transwell collagen coated Nuclepore filters. Two min NMN or TEA unidirectional transepithelial flux and simultaneous cellular uptake were measured. Transport of NMN and TEA from basolateral to apical side was temperature-dependent, saturable and competitively inhibited by each other or by mepiperphenidol. NMN and TEA transport from the apical to the basolateral side was very slow, only slightly faster than that of mannitol. Apparent kinetic parameters of basolateral to apical transcellular flux were measured. For NMN apparent Km = 133.6 +/- 35.4 microM, Vmax = 48.3 +/- 5.6 pmol/cm2.2 min. For TEA apparent Km = 11.4 +/- 2.2 microM, Vmax = 42.1 +/- 1.8 pmol/cm2.2 min. Kinetic parameters of cellular uptake were also estimated. For NMN apparent Km = 436.9 +/- 139.8 microM, Vmax = 143.7 +/- 30.5 pmol/micrograms DNA. For TEA apparent Km = 50.3 +/- 7.2 microM, Vmax = 26.5 +/- 2.5 pmol/micrograms DNA. It is concluded that LLC-PK1 cells transport NMN and TEA from the basolateral to the apical side; this flux corresponds to the secretory transport of the renal proximal tubule. NMN and TEA share the same transport system, but NMN has a lower affinity.