Determination of topological structure of ARL6ip1 in cells: identification of the essential binding region of ARL6ip1 for conophylline

Abstract

Conophylline (CNP) has various biological activities, such as insulin production. A recent study identified ADP-ribosylation factor-like 6-interacting protein 1 (ARL6ip1) as a direct target protein of CNP. In this study, we revealed that ARL6ip1 is a three-spanning transmembrane protein and determined the CNP-binding domain of ARL6ip1 by deletion mutation analysis of ARL6ip1 with biotinyl-amino-CNP. These results suggest that CNP is expected to be useful for future investigation of ARL6ip1 function in cells. Because of the anti-apoptotic function of ARL6ip1, CNP may be an effective therapeutic drug and/or a novel chemosensitizer for human cancers and other diseases.

Keywords: ADP-ribosylation factor-like 6-interacting protein 1; ARL6ip1; BCNP; CNP; Conophylline; ER; Gaussia luciferase; Gluc; Redox-sensitive luciferase assay; TM; Topology; biotinyl-aminoconophylline; conophylline; endoplasmic reticulum; transmembrane.