Pharmacological investigation of the role of aplnr and its post-translational modification

Abstract

Apelin receptor, Aplnr, is a member of G protein coupled receptor. The amino acid sequence suggests that Aplnr has two N-glycosylation sites. Here, we have detected the Aplnr proteins which was overexpressed in HEK293 cells, or was purified from the central nervous system of the mouse. Western blot analysis revealed that, in the HEK cells that was overexpressed of Aplnr, there are two bands on the membrane around the molecular weight of Aplnr, that have already been reported. Among of the two, the band with slow mobility, was disappeared by the sample incubation with Peptide-N-Glycosidase F (PNGase F) for cleaving N-glycosylation sites. On the other hand, when attempting to detect Aplnr using the mouse spinal cord, multiple bands at approximately 20 kDa greater than the expected molecular weights were detected. Among of them, the band with a slow mobility, was disappeared by PNGase F in the same manner as described above. Therefore, it was suggested that Aplnr in the central nervous system expresses with several post-translational modification, including N-glycosylation, and it functions differently to peripheral organs.