A lipid scramblase TMEM41B is involved in the processing and transport of GPI-anchored proteins

Abstract

Protein modification by glycosylphosphatidylinositol (GPI) takes place in the endoplasmic reticulum (ER). GPI-anchored proteins (GPI-APs) formed in the ER are transported to the cell surface through the Golgi apparatus. During transport, the GPI-anchor structure is processed. In most cells, an acyl chain modified to the inositol of GPI is removed by a GPI-inositol deacylase, PGAP1, in the ER. Inositol-deacylated GPI-APs become sensitive to bacterial phosphatidylinositol-specific phospholipase C (PI-PLC). We previously reported that GPI-APs are partially resistant to PI-PLC when PGAP1 activity is weakened by the deletion of selenoprotein T (SELT) or cleft lip and palate transmembrane protein 1 (CLPTM1). In this study, we found that the loss of TMEM41B, an ER-localized lipid scramblase, restored PI-PLC sensitivity of GPI-APs in SELT-knockout (KO) and CLPTM1-KO cells. In TMEM41B-KO cells, the transport of GPI-APs as well as transmembrane proteins from the ER to the Golgi was delayed. Furthermore, the turnover of PGAP1, which is mediated by ER-associated degradation, was slowed in TMEM41B-KO cells. Taken together, these findings indicate that inhibition of TMEM41B-dependent lipid scrambling promotes GPI-AP processing in the ER through PGAP1 stabilization and slowed protein trafficking.

Keywords: TMEM41B; glycosylphosphatidylinositol. Abbreviations: Baf A1, bafilomycin A1; BFP, blue fluorescent protein; CHX, cycloheximide; CLPTM1, cleft lip and palate transmembrane protein 1; COP-II, coat protein complex II; DKO, double knockout; EtNP, ethanolamine-phosphate; ER, endoplasmic reticulum; ERAD, ER-associated degradation; GPI, glycosylphosphatidylinositol; GPI-AP, GPI-anchored protein; KO, knockout; Man, mannose; PI, phosphatidylinositol; PI-PLC, phosphatidylinositol-specific phospholipase C; SELT, selenoprotein T; sgRNA, single-guide RNA; lipid remodeling; protein trafficking; scramblase.