Human apolipoprotein C-I accounts for the ability of plasma high density lipoproteins to inhibit the cholesteryl ester transfer protein activity.
The aim of the present study was to identify the protein that accounts for the cholesteryl ester transfer protein (CETP)-inhibitory activity that is specifically associated with human plasma high density lipoproteins (HDL). To this end, human HDL apolipoproteins were fractionated by preparative polyacrylamide gradient gel electrophoresis, and 30 distinct protein fractions with molecular masses ranging from 80 down to 2 kDa were tested for their ability to inhibit CETP activity. One single apolipoprotein fraction was able to completely inhibit CETP activity. The N-terminal sequence of the 6-kDa protein inhibitor matched the N-terminal sequence of human apoC-I, the inhibition was completely blocked by specific anti-apolipoprotein C-I antibodies, and mass spectrometry analysis confirmed the identity of the isolated inhibitor with full-length human apoC-I. Pure apoC-I was able to abolish CETP activity in a concentration-dependent manner and with a high efficiency (IC(50) = 100 nmol/liter). The inhibitory potency of total delipidated HDL apolipoproteins completely disappeared after a treatment with anti-apolipoprotein C-I antibodies, and the apoC-I deprivation of native plasma HDL by immunoaffinity chromatography produced a mean 43% rise in cholesteryl ester transfer rates. The main localization of apoC-I in HDL and not in low density lipoprotein in normolipidemic plasma provides further support for the specific property of HDL in inhibiting CETP activity.
Gautier
T
T
Laboratoire de Biochimie des Lipoprotéines-INSERM U498, Hôpital du Bocage, BP1542, 21034 Dijon Cedex, France.
Masson
D
D
de Barros
J P
JP
Athias
A
A
Gambert
P
P
Aunis
D
D
Metz-Boutigue
M H
MH
Lagrost
L
L
D000595:Amino Acid Sequence; D053302:Apolipoprotein C-I; D001056:Apolipoproteins C / Q000502:physiology*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053480:Cholesterol Ester Transfer Proteins; D008076:Cholesterol, HDL / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D008078:Cholesterol, LDL / Q000502:physiology; D006023:Glycoproteins; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding
D053302:Apolipoprotein C-I; D001056:Apolipoproteins C; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D008076:Cholesterol, HDL; D008078:Cholesterol, LDL; D006023:Glycoproteins