Characterization of human lysophospholipid acyltransferase 3.
Esterifying lysophospholipids may serve a variety of functions, including phospholipid remodeling and limiting the abundance of bioactive lipids. Recently, a yeast enzyme, Lpt1p, that esterifies an array of lysophospholipids was identified. Described here is the characterization of a human homolog of LPT1 that we have called lysophosphatidylcholine acyltransferase 3 (LPCAT3). Expression of LPCAT3 in Sf9 insect cells conferred robust esterification of lysophosphatidylcholine in vitro. Kinetic analysis found apparent cooperativity with a saturated acyl-CoA having the lowest K0.5 (5 microM), a monounsaturated acyl-CoA having the highest apparent Vmax (759 nmol/min/mg), and two polyunsaturated acyl-CoAs showing intermediate values. Lysophosphatidylethanolamine and lysophosphatidylserine were also utilized as substrates. Electrospray ionization mass spectrometric analysis of phospholipids in Sf9 cells expressing LPCAT3 showed a relative increase in phosphatidylcholine containing saturated acyl chains and a decrease in phosphatidylcholine containing unsaturated acyl chains. Targeted reduction of LPCAT3 expression in HEK293 cells had essentially an opposite effect, resulting in decreased abundance of saturated phospholipid species and more unsaturated species. Reduced LPCAT3 expression resulted in more apoptosis and distinctly fewer lamellipodia, suggesting a necessary role for lysophospholipid esterification in maintaining cellular function and structure.
Department of Biology, Drexel University, Philadelphia, PA 19104, USA.
Shilpa
S
Jain
Xiaoling
X
Zhang
Preeti J
PJ
Khandelwal
Aleister J
AJ
Saunders
Brian S
BS
Cummings
Peter
P
Oelkers
D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000235:genetics / Q000378:metabolism*; D000214:Acyl Coenzyme A / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002478:Cells, Cultured; D005227:Fatty Acids / Q000032:analysis*; D006801:Humans; D007700:Kinetics; D008246:Lysophospholipids / Q000378:metabolism*; D010743:Phospholipids / Q000032:analysis*; D011554:Pseudopodia / Q000648:ultrastructure; D034622:RNA Interference; D013379:Substrate Specificity; D014162:Transfection
D000214:Acyl Coenzyme A; D005227:Fatty Acids; D008246:Lysophospholipids; D010743:Phospholipids; D008243:1-Acylglycerophosphocholine O-Acyltransferase