Diacylglycerol is the preferred substrate in high density lipoproteins for human hepatic lipase.
The hydrolysis of HDL phospholipids (PL) and glycerides by hepatic lipase (HL) has been investigated in native and reconstituted HDL particles (Lp2A-I). Fasting, normolipidemic HDL exhibit total lipid hydrolytic rates of between 10 and 36 nM FA/h per microM PL. Of the total fatty acids liberated with HDL3 only 1% are from triolein (TG), while 49% are from diolein (DG) and 50% are from PL. A spherical reconstituted particle containing 2 molecules of apoA-I, 120 molecules of PL, and 20 molecules of TG exhibits a total lipid hydrolytic rate of 18 nM FA/h per microM PL and 93% of the fatty acids liberated are from PL. Inclusion of 40 molecules of TG into the Lp2A-I particle doubles the rate of fatty acid hydrolysis by HL through a stimulation of TG hydrolysis. Further addition of 10 molecules of DG to the Lp2A-I complex has no effect on the overall rates of hydrolysis, but changes the substrate specificity, wherein 61% of the fatty acids are from DG and both TG and PL hydrolytic rates are significantly reduced. Increasing the amount of DG in the Lp2A-I particle further stimulates total lipid hydrolysis by raising DG hydrolytic rates at the expense of PL and TG hydrolysis. A particle containing 10 molecules of TG and 40 molecules DG yields the fastest lipid hydrolytic rate of 143 nM FA/h per microM PL, which constitutes 96% DG hydrolysis, 3% TG hydrolysis, and 1% PC hydrolysis. These data indicate that hepatic lipase acts primarily as a surface lipid lipase with HDL particles. DG is the preferred substrate of HL in HDL and the HDL-DG content regulates the hydrolysis of both PL and TG by HL.
Coffill
C R
CR
University of Ottawa Heart Institute, and Department of Biochemistry, University of Ottawa, Ontario, Canada.
Ramsamy
T A
TA
Hutt
D M
DM
Schultz
J R
JR
Sparks
D L
DL
D002250:Carbon Radioisotopes; D004075:Diglycerides / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D010316:Particle Size; D010713:Phosphatidylcholines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D013379:Substrate Specificity; D014304:Triolein / Q000378:metabolism; D014316:Tritium
D002250:Carbon Radioisotopes; D004075:Diglycerides; D008075:Lipoproteins, HDL; D010713:Phosphatidylcholines; D010743:Phospholipids; D014316:Tritium; D014304:Triolein; D008049:Lipase; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; C013965:diolein