title: Functional and structural properties of lipid-associated apolipoprotein J (clusterin).
abstract: Apolipoprotein J (apoJ, clusterin) is a multifunctional protein normally associated with lipids in plasma and cerebrospinal fluid, and secreted as lipoparticles by hepatocytes and astrocytes. To investigate whether the structural and functional properties of apoJ are modulated upon binding to lipids, we prepared apoJ high-density lipoprotein (HDL)-like particles employing either synthetic or plasma HDL-derived lipids. The majority of the resulting lipoparticles contained one molecule of apoJ per particle and exhibited the same alpha2 electrophoretic mobility characteristic of apoJ-containing plasma HDL. Particle size seemed to be dependent on the presence of cholesterol in the lipid mixture and ranged from diameters of 10 nm in the presence of cholesterol to 20 nm in the absence of cholesterol. CD analysis and Fourier-transform infrared spectroscopy revealed similar changes in the apoJ secondary structure induced by its incorporation into lipoparticles, namely a decrease in alpha-helix content and an increase in beta-turn structures. Two functional assays, the binding interaction with Alzheimer's amyloid beta peptides and the inhibitory activity of the complement membrane-attack complex, did not detect any changes in apoJ activity following its lipidation (P>0.05). On the contrary, the binding affinity to the cellular receptor megalin was enhanced significantly (P<0.01) after the association with lipids; the K(d) value decreased from 78.8+/-10.7 nM for the delipidated form to 37. 0+/-7.3 nM for apoJ-HDL. Although it is not known whether the structural changes observed are directly responsible for the higher receptor-binding affinity, the data suggest that the complement inhibition and amyloid beta-binding motifs are located in areas of the molecule different from those involved in the apoJ-megalin interaction.
authors.affiliation: Department of Pathology, New York University School of Medicine, 550 First Avenue, New York, NY 10016, USA.
authors.forename: M
authors.initials: M
authors.lastname: Calero
authors.forename: T
authors.initials: T
authors.lastname: Tokuda
authors.forename: A
authors.initials: A
authors.lastname: Rostagno
authors.forename: A
authors.initials: A
authors.lastname: Kumar
authors.forename: B
authors.initials: B
authors.lastname: Zlokovic
authors.forename: B
authors.initials: B
authors.lastname: Frangione
authors.forename: J
authors.initials: J
authors.lastname: Ghiso
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism; D000818:Animals; D001053:Apolipoproteins / Q000097:blood / Q000737:chemistry* / Q000648:ultrastructure; D051152:Clusterin; D003167:Complement Activation; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D003432:Cross-Linking Reagents; D006023:Glycoproteins / Q000097:blood / Q000737:chemistry* / Q000648:ultrastructure; D006801:Humans; D008075:Lipoproteins, HDL / Q000737:chemistry* / Q000648:ultrastructure; D051379:Mice; D018832:Molecular Chaperones; D017433:Protein Structure, Secondary; D018110:Receptors, Lipoprotein / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D001053:Apolipoproteins; C495242:CLU protein, human; C495243:Clu protein, mouse; D051152:Clusterin; D015938:Complement Membrane Attack Complex; D003432:Cross-Linking Reagents; D006023:Glycoproteins; D008075:Lipoproteins, HDL; D018832:Molecular Chaperones; D018110:Receptors, Lipoprotein