title: Synthesis and secretion of wild-type and mutant human plasma cholesteryl ester transfer protein in baculovirus-transfected insect cells: the carboxyl-terminal region is required for both lipoprotein binding and catalysis of transfer.
abstract: Functional plasma cholesteryl ester transfer protein (CETP; 476 amino acids) has been expressed in baculovirus-transfected Sf9 insect cells by using a full-length cDNA derived from a human placental library. The product bound to each major plasma lipoprotein class, and it catalyzed the transfer of both cholesteryl esters and triglyceride. CETP species with overlapping deletions were generated in the carboxyl-terminal region. These mutants were defective in cholesteryl ester and triglyceride transfer. Structural and functional analysis suggests that normal lipoprotein binding and effective catalysis may require the carboxyl-terminal sequence -Phe-Leu-Leu-Leu- (residues 454-457), possibly with the involvement of other sequences in the carboxyl-terminal region. A similar sequence is contained in several other proteins whose functions involve binding nonpolar lipids, including lecithin: cholesterol acyltransferase, lipopolysaccharide-binding protein, bactericidal permeability-increasing protein, cholesterol 7 alpha-hydroxylase, cholesterol esterase, and hormone-sensitive lipase. These data suggest that a conserved neutral lipid-binding sequence may be one important factor in the activity of CETP and possibly in several other proteins of plasma and cellular lipid metabolism.
authors.affiliation: Cardiovascular Research Institute, University of California Medical Center, San Francisco 94143.
authors.forename: J
authors.initials: J
authors.lastname: Au-Young
authors.forename: C J
authors.initials: CJ
authors.lastname: Fielding
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000378:metabolism*; D006023:Glycoproteins; D006801:Humans; D066298:In Vitro Techniques; D008074:Lipoproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D006023:Glycoproteins; D008074:Lipoproteins; D011994:Recombinant Proteins