HK-2 Human Renal Proximal Tubule Cells as a Model for G Protein–Coupled Receptor Kinase Type 4–Mediated Dopamine 1 Receptor Uncoupling

HK-2 human renal proximal tubule cells (RPTC) are commonly used in the in vitro study of “normal” RPTCs. We discovered recently that HK-2 cells are uncoupled from dopamine 1 receptor (D 1 R) adenylyl cyclase (AC) stimulation. We hypothesized that G protein–coupled receptor kinase type 4 (GRK4) single nucleotide polymorphisms may be responsible for the D 1 R/AC uncoupling in HK-2. This hypothesis was tested by genotyping GRK4 single nucleotide polymorphisms, measuring D 1 -like receptor agonist (fenoldopam)-stimulated cAMP accumulation, quantifying D 1 R inhibition of sodium transport, and testing the ability of GRK4 small interfering RNA to reverse the D 1 R/AC uncoupling. We compared HK-2 with 2 normally coupled human RPTC cell lines and 2 uncoupled RPTC cell lines. The HK-2 cell line was found to have 4 of 6 potential GRK4 single nucleotide polymorphisms known to uncouple the D 1 R from AC (namely, R65L, A142V, and A486V). AC response to fenoldopam stimulation was increased in the 2 normally coupled human RPTC cell lines (FEN: 2.02±0.05-fold and 2.33±0.19-fold over control; P P