Mechanism of cholesteryl ester transfer protein inhibition by a neutralizing monoclonal antibody and mapping of the monoclonal antibody epitope.
The plasma cholesteryl ester transfer protein (CETP, Mr 74,000) has a binding site for neutral lipid which can readily equilibrate with lipoprotein cholesteryl esters or triglycerides. Recently, a monoclonal antibody (TP2) was obtained which neutralizes the cholesteryl ester (CE) and triglyceride (TG) transfer activities of the CETP. In this report, the epitope of the inhibitory monoclonal antibody has been localized to a hydrophobic 26-amino acid sequence at the COOH terminus of CETP. The Fab fragments of TP2 caused partial (50%) inhibition of CE transfer and complete inhibition of TG transfer by the CETP. Similarly, the Fab fragments inhibited (37%) the binding of CE to the CETP and abolished the binding of TG to the CETP. Surprisingly, the TP2 Fab was also found to enhance the binding of CETP to plasma lipoproteins and to phospholipid vesicles. In conclusion, the TP2 monoclonal antibody inhibits lipid transfer by blocking the uptake of lipid by CETP. The COOH-terminal epitope may be in or near the neutral lipid binding site. Occupancy of this site by TP2 Fab fragments or by neutral lipid may result in a conformational change of CETP causing enhanced binding to lipoproteins or vesicles.
Swenson
T L
TL
Department of Medicine, Columbia University College of Physicians and Surgeons, New York 10032.
Hesler
C B
CB
Brown
M L
ML
Quinet
E
E
Trotta
P P
PP
Haslanger
M F
MF
Gaeta
F C
FC
Marcel
Y L
YL
Milne
R W
RW
Tall
A R
AR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology*; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000378:metabolism*; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006023:Glycoproteins; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000502:physiology; D050356:Lipid Metabolism; D008074:Lipoproteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010449:Peptide Mapping
D000911:Antibodies, Monoclonal; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D000939:Epitopes; D006023:Glycoproteins; D007140:Immunoglobulin Fab Fragments; D008074:Lipoproteins