MOGAT2 contains a phosphate acyltransferase-like domain and 2 predicted N-linked glycosylation sites. Northern blot analysis revealed expression of 7.2-, 3.0-, and 1.3-kb transcripts in liver, stomach, and small intestine, with lower expression in colon and kidney. A probe specific to the truncated splice variant hybridized mainly to the 1.3-kb transcript in stomach. Immunofluorescence analysis of transiently transfected COS-7 cells showed that MGAT2 displayed a pattern consistent with localization in the endoplasmic reticulum.Substrate specificity assays of MOGAT2 demonstrated that MOGAT2 can catalyze the acylation of rac-1- and sn-2-monooleoylglycerols. MOGAT2 uses a broad range of fatty acyl-CoAs but prefers monoacylglycerols containing unsaturated fatty acids.