title: New insights into the functions of PIGF, a protein involved in the ethanolamine phosphate transfer steps of glycosylphosphatidylinositol biosynthesis.
abstract: PIGF is a protein involved in the ethanolamine phosphate (EtNP) transfer steps of glycosylphosphatidylinositol (GPI) biosynthesis. PIGF forms a heterodimer with either PIGG or PIGO, two enzymes that transfer an EtNP to the second or third mannoses of GPI respectively. Heterodimer formation is essential for stable and regulated expression of PIGO and PIGG, but the functional significance of PIGF remains obscure. In the present study, we show that PIGF binds to PIGO and PIGG through distinct molecular domains. Strikingly, C-terminal half of PIGF was sufficient for its binding to PIGO and PIGG and yet this truncation mutant could not complement the PIGF defective mutant cells, suggesting that heterodimer formation is not sufficient for PIGF function. Furthermore, we identified a highly conserved motif in PIGF and demonstrated that the motif is not involved in binding to PIGO or PIGG, but critical for its function. Finally, we identified a PIGF homologue from Trypanosoma brucei and showed that it binds specifically to the T. brucei PIGO homologue. These data together support the notion that PIGF plays a critical and evolutionary conserved role in the ethanolamine-phosphate transfer-step, which cannot be explained by its previously ascribed binding/stabilizing function. Potential roles of PIGF in GPI biosynthesis are discussed.
authors: *Laboratory of Immunoglycobiology, WPI Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
authors: Matthew J
authors: MJ
authors: Stokes
authors: Yoshiko
authors: Y
authors: Murakami
authors: Yusuke
authors: Y
authors: Maeda
authors: Taroh
authors: T
authors: Kinoshita
authors: Yasu S
authors: YS
authors: Morita
mesh_terms: D000818:Animals; D002460:Cell Line; D004982:Ethanolaminephosphotransferase / Q000235:genetics / Q000378:metabolism; D004983:Ethanolamines / Q000378:metabolism*; D019143:Evolution, Molecular; D017261:Glycosylphosphatidylinositols / Q000096:biosynthesis*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014346:Trypanosoma brucei brucei / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D004983:Ethanolamines; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; C098069:PIGF protein, human; C005448:phosphorylethanolamine; D004982:Ethanolaminephosphotransferase