The c‐terminal region of BLT2 restricts its localization to the lateral membrane in a LIN7C‐dependent manner

Abstract

Leukotriene B₄ receptor type 2 (BLT2) is a G protein‐coupled receptor (GPCR) mainly expressed in epithelial cells, where it enhances barrier function. A unique characteristic of BLT2 is its restricted localization to the lateral membrane. However, the molecular mechanism underlying the localization of BLT2 to the lateral membrane and the physiological roles of laterally localized BLT2 are unknown. BLT1 is the most homologous GPCR to BLT2 and localizes to both the apical and lateral membranes. In this study, we generated chimeric receptors of BLT2 and BLT1 as well as deletion mutants of BLT2 to determine the region(s) of BLT2 responsible for its localization. Chimeric receptors containing the C‐terminal domain of BLT2 localized only to the lateral membrane, and the C‐terminal deletion mutant of BLT2 accumulated at the Golgi apparatus. Furthermore, the middle and C‐terminal regions of BLT2 were important for maintaining epithelial barrier function. Proteomics analysis using the chimeric BLT‐ascorbate peroxidase 2 biotinylation method showed that some proteins involved in intracellular protein transport, cell‐cell junctions, and actin filament binding were located very close to the C‐terminal domain of BLT2. Knockdown of lin‐7 homolog C (LIN7C), a membrane trafficking protein, led to accumulation of BLT2 in the Golgi apparatus, resulting in diminished epithelial barrier function. These results suggest that the C‐terminal region of BLT2 plays an important role in the transport of BLT2 from the Golgi apparatus to the plasma membrane in a LIN7C‐dependent manner.