[HTML][HTML] Protein kinase C (PKC)-induced phosphorylation of ROMK1 is essential for the surface expression of ROMK1 channels

DH Lin, H Sterling, KM Lerea, G Giebisch… - Journal of Biological …, 2002 - ASBMB
We carried out in vitrophosphorylation assays to determine whether ROMK1 is a substrate of
protein kinase C (PKC) and used the two-electrode voltage clamp method to investigate the
role of serine residues 4, 183, and 201, the three putative PKC phosphorylation sites, in the
regulation of ROMK1 channel activity. Incubation of the purified His-tagged ROMK1 protein
with PKC and radiolabeled ATP resulted in 32 P incorporation into ROMK1 detected by
autoradiography. Moreover, the in vitro phosphorylation study of three synthesized peptides …