Characterization of chicken cystatin binding to rat renal brush-border membranes

Abstract

Chicken cystatin, a homologue of human cystatin C, like other low-molecular-weight proteins is metabolized by renal proximal tubule cells. However, the precise mechanism(s) of this process has not been elucidated yet. To characterize chicken cystatin binding to renal brush-border membranes, the incubation of fluorescein labelled protein with rat cortical homogenate was performed. Saturation-dependent and reversible binding with low affinity ( K d = 3.67–4.07 μM) and high capacity ( B max = 2.32–2.79 nmol/mg) was observed. Bovine albumin was the most potent competitor ( K i = 0.7 μM) among other megalin/cubilin ligands tested. The presence of Ca + 2 ions was necessary to effective cystatin binding by brush-border membranes. Obtained data strongly support the hypothesis that chicken cystatin is a novel ligand for megalin/cubilin receptors tandem on proximal tubular cells.