Tubular inhibition destroys charge selectivity for anionic and neutral horseradish peroxidase

The fractional clearance of [3H]anionic HRP and [3H]neutral HRP in the isolated perfused kidney as determined by radioactivity analysis was 0.0160+/-0.0028 (n=6) and 0.0388+/-0.0076 (n=8) respectively. The apparent charge selectivity for both the anionic and neutral forms of HRP observed was destroyed with the inclusion of the tubular uptake inhibitors, 150 mM lysine and 10 mM NH4Cl, in the perfusate. In the presence of 150 mM lysine, the clearances of [3H]anionic HRP and [3H]neutral HRP were 0.0645+/-0.0110 (n=4) and 0. 0784+/-0.0120 (n=4) respectively, and 0.0564+/-0.0035 (n=4) and 0. 0694+/-0.0054 (n=4) respectively in the presence of 10 mM NH4Cl. The clearance for both the anionic and neutral HRP probes in these tubular uptake inhibited systems fits precisely the size selective characteristics of the glomerular capillary wall as determined by transport probes calibrated for hydrodynamic size by size exclusion chromatography. The tubular uptake inhibitors were observed not to alter glomerular permselectivity as determined using polydisperse dextran fractions and the behaviour of neutral HRP. This study demonstrates that charge selectivity for differently charged proteins is not as great as originally thought and suggests that the clearance differences between anionic and neutral forms may be due to differential handling by the tubules.