Identification and characterization of a gene encoding human LPGAT1, an endoplasmic reticulum-associated lysophosphatidylglycerol acyltransferase.
Phosphatidylglycerol (PG) is an important membrane polyglycerolphospholipid required for the activity of a variety of enzymes and is a precursor for synthesis of cardiolipin and bis(monoacylglycerol) phosphate. PG is subjected to remodeling subsequent to its de novo biosynthesis to incorporate appropriate acyl content for its biological functions and to prevent the harmful effect of lysophosphatidylglycerol (LPG) accumulation. The enzymes involved in the remodeling process have not yet been identified. We report here the identification and characterization of a human gene encoding an acyl-CoA: lysophosphatidylglycerol acyltransferase (LPGAT1). Expression of the LPGAT1 cDNA in Sf9 insect and COS-7 cells led to a significant increase in LPG acyltransferase activity. In contrast, no significant acyltransferase activities were detected against glycerol 3-phosphate or a variety of lysophospholipids, including lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylinositol, and lysophosphatidylserine. The recombinant human LPGAT1 enzyme recognized various acyl-CoAs and LPGs as substrates but demonstrated clear preference to long chain saturated fatty acyl-CoAs and oleoyl-CoA as acyl donors, which is consistent with the lipid composition of endogenous PGs identified from different tissues. Kinetic analyses of LPGAT1 expressed in COS-7 cells showed that oleoyl-LPG was preferred over palmitoyl-LPG as an acyl receptor, whereas oleoyl-CoA was preferred over lauroyl-CoA as an acyl donor. Consistent with its proposed microsomal origin, LPGAT1 was localized to the endoplasmic reticulum by subcellular fractionation and immunohistochemical analyses. Northern blot analysis indicated that the human LPGAT1 was widely distributed, suggesting a dynamic functional role of the enzyme in different tissues.
Yang
Yanzhu
Y
Endocrine Research, Lilly Research Laboratories, Eli Lilly and Co., Indianapolis, IN 46285, USA.
Cao
Jingsong
J
Shi
Yuguang
Y
D000217:Acyltransferases / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D015152:Blotting, Northern; D015153:Blotting, Western; D019556:COS Cells; D002308:Cardiolipins / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005994:Glycerophosphates / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D007313:Insecta; D007700:Kinetics; D008246:Lysophospholipids / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008861:Microsomes / Q000378:metabolism; D008954:Models, Biological; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010101:Oxygen Consumption; D010455:Peptides / Q000737:chemistry; D010712:Phosphatidic Acids / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D002308:Cardiolipins; D018076:DNA, Complementary; D005994:Glycerophosphates; D008246:Lysophospholipids; D010455:Peptides; D010712:Phosphatidic Acids; D011994:Recombinant Proteins; C025059:lysophosphatidylserine; C029620:alpha-glycerophosphoric acid; D000217:Acyltransferases; C496548:LPGAT1 protein, human