The in vitro formation of HDL2 during the action of LCAT: the role of triglyceride-rich lipoproteins.
We examined the effects of lecithin:cholesterol acyl transferase (LCAT) and of lipoprotein lipase (LPL) on the conversion of high density lipoproteins (HDL) towards fractions of lower densities using the analytical ultracentrifuge. Freshly isolated whole plasma was incubated for 24 h at 37 degrees C in the presence or absence of active enzyme systems. In some cases, lipoproteins were removed by selective precipitations; alternatively, we added triglyceride-rich lipoproteins (TGRLP) or Intralipid to the incubations. The results are as follows. 1) The incubation of whole plasma containing active LCAT leads to a conversion of HDL3 to a fraction of lower density, notably HDL2a. If LCAT is inhibited, the conversion is far less pronounced. 2) If very low and low density lipoproteins are removed by phosphotungstate precipitation and the supernatant is incubated with LCAT, HDL3 shifts towards higher densities. 3) The presence of phosphatidylcholine/cholesterol liposomes or the presence of blood cells as a source of additional LCAT substrate had only little influence on the HDL conversion in our system. 4) The addition of TGRLP or of Intralipid at minimal ratios of 2.5:1 caused an almost complete conversion of HDL3 to HDL2b. This conversion was dependent on active LCAT. 5) LPL also caused a shift of HDL3 to HDL2a if TGRLP was present. HDL2b, however, was not formed by LPL unless LCAT was active.
D002784:Cholesterol / Q000097:blood; D004589:Electrophoresis, Disc; D005215:Fasting; D005217:Fat Emulsions, Intravenous; D006801:Humans; D066298:In Vitro Techniques; D008071:Lipoprotein Lipase / Q000378:metabolism; D008074:Lipoproteins / Q000097:blood*; D008075:Lipoproteins, HDL / Q000096:biosynthesis* / Q000097:blood; D053439:Lipoproteins, HDL2; D008081:Liposomes; D010316:Particle Size; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000097:blood; D014280:Triglycerides / Q000097:blood*; D014461:Ultracentrifugation
D005217:Fat Emulsions, Intravenous; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D053439:Lipoproteins, HDL2; D008081:Liposomes; D010713:Phosphatidylcholines; D014280:Triglycerides; D002784:Cholesterol; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D008071:Lipoprotein Lipase