title: Identification of small subunit of serine palmitoyltransferase a as a lysophosphatidylinositol acyltransferase 1-interacting protein.
abstract: Lysophosphatidylinositol acyltransferase 1 (LPIAT1), also known as MBOAT7, is a phospholipid acyltransferase that selectively incorporates arachidonic acid (AA) into the sn-2 position of phosphatidylinositol (PI). We previously demonstrated that LPIAT1 regulates AA content in PI and plays a crucial role in brain development in mice. However, how LPIAT1 is regulated and which proteins function cooperatively with LPIAT1 are unknown. In this study, using a split-ubiquitin membrane yeast two-hybrid system, we identified the small subunit of serine palmitoyltransferase a (ssSPTa) as an LPIAT1-interacting protein. ssSPTa co-immunoprecipitated and colocalized with LPIAT1 in cultured mammalian cells. Knockdown of ssSPTa decreased the LPIAT1-dependent incorporation of exogenous AA into PI but did not affect the in vitro enzyme activity of LPIAT1 in the microsomal fraction. Interestingly, knockdown of ssSPTa decreased the protein level of LPIAT1 in the crude mitochondrial fraction but not in total homogenate or the microsomal fraction. LPIAT1 was localized to the mitochondria-associated membrane (MAM), where AA-selective acyl-CoA synthetase is enriched. These results suggest that ssSPTa plays a role in fatty acid remodeling of PI, probably by facilitating the MAM localization of LPIAT1.
authors.affiliation: Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, 113-0033, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Hirata
authors.forename: Natsumi
authors.initials: N
authors.lastname: Yamamori
authors.forename: Nozomu
authors.initials: N
authors.lastname: Kono
authors.forename: Hyeon-Cheol
authors.initials: HC
authors.lastname: Lee
authors.forename: Takao
authors.initials: T
authors.lastname: Inoue
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Arai
mesh_terms: D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D051336:Mitochondrial Membranes / Q000201:enzymology; D010716:Phosphatidylinositols / Q000378:metabolism; D011485:Protein Binding; D021122:Protein Subunits / Q000378:metabolism*; D021381:Protein Transport; D051102:Serine C-Palmitoyltransferase / Q000378:metabolism*; D013107:Sphingolipids / Q000096:biosynthesis; D034421:Sus scrofa
chemical_list: D010716:Phosphatidylinositols; D021122:Protein Subunits; D013107:Sphingolipids; D016718:Arachidonic Acid; D000217:Acyltransferases; C021522:lysophosphatidylinositol acyltransferase; D051102:Serine C-Palmitoyltransferase; C541445:ssSPTa