Use of Intralipid for kinetic analysis of HDL apoC-III: evidence for a homogeneous kinetic pool of apoC-III in plasma.
Apolipoprotein C-III (apoC-III) is an important regulator of lipoprotein metabolism. Radioisotope and stable isotope kinetic studies show differing results in relation to the kinetics of apoC-III in HDL. Kinetic analysis of HDL apoC-III may be difficult because of its low concentration, as well as the presence of other apoproteins at higher concentration, in the HDL fraction. We used Intralipid(R) (IL), known to preferentially extract apoC proteins from plasma, as a means of extracting apoC-III from HDL before apoprotein separation by isoelectric focusing gel electrophoresis for the measurement of tracer enrichment. Protein purity was assessed by an isoleucine-to-leucine (Ile/Leu) ratio, as apoC-III contains no isoleucine. We compared apoC-III kinetics in 14 men using a bolus infusion of deuterated leucine. The Ile/Leu ratio for IL-extracted HDL (IL-HDL) apoC-III (3.0 +/- 0.7%) was not different from that of VLDL apoC-III (2.6 +/- 0.6%) but was significantly lower than that of untreated HDL apoC-III (9.0 +/- 2.9%) (P < 0.001). The isotopic enrichment curves and fractional catabolic rates (FCRs) for IL-HDL apoC-III were not different from those of VLDL apoC-III. In contrast, HDL apoC-III had significantly lower isotopic enrichments and FCRs than IL-HDL apoC-III (P < 0.001). In conclusion, this simple IL method can be used to isolate apoC-III from HDL with minimal interference from other HDL apoproteins, and it demonstrates that the kinetics of apoC-III in VLDL and HDL are similar, supporting the concept of a single kinetically homogeneous pool of apoC-III in plasma.
Metabolic Research Centre, School of Medicine and Pharmacology, University of Western Australia, Perth, Western Australia.
Minh N
MN
Nguyen
Dick C
DC
Chan
Kevin P
KP
Dwyer
Paul
P
Bolitho
Gerald F
GF
Watts
P Hugh R
PH
Barrett
D053305:Apolipoprotein C-III; D001056:Apolipoproteins C / Q000097:blood* / Q000378:metabolism; D005217:Fat Emulsions, Intravenous / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D007532:Isoleucine / Q000032:analysis; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000097:blood* / Q000378:metabolism; D008079:Lipoproteins, VLDL / Q000097:blood / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008954:Models, Biological
D053305:Apolipoprotein C-III; D001056:Apolipoproteins C; D005217:Fat Emulsions, Intravenous; D008075:Lipoproteins, HDL; D008079:Lipoproteins, VLDL; D007532:Isoleucine