title: Adduction of cholesterol 5,6-secosterol aldehyde to membrane-bound myelin basic protein exposes an immunodominant epitope.
abstract: Myelin degradation in the central nervous system (CNS) is a clinical hallmark of multiple sclerosis (MS). A reduction in the net positive charge of myelin basic protein (MBP) via deimination of arginine to citrulline has been shown to correlate strongly with disease severity and has been linked to myelin instability and a defect that precedes neurodegeneration and leads to autoimmune attack. Recently, we have shown that lipid-derived aldehydes, such as cholesterol 5,6-secosterols atheronal A (1a) and atheronal B (1b), modulate the misfolding of certain proteins such as apolipoprotein B(100), β-amyloid, α-synuclein, and κ- and λ-antibody light chains in a process involving adduction of the hydrophobic aldehyde to lysine side chains, resulting in a decrease in the net positive charge of the protein. In this study, we show that the presence of either atheronal A (1a) or atheronal B (1b) in large unilamellar vesicles (cyt-LUVs) with the lipid composition found in the cytosolic myelin sheath and bovine MBP (bMBP) leads to an atheronal concentration-dependent increase in the surface exposure of the immunodominant epitope (V86-T98) as determined by antibody binding. Other structural changes in bMBP were also observed; specifically, 1a and 1b induce a decrease in the surface exposure of L36-P50 relative to control cyt-LUVs as measured both by antibody binding and by a reduction in the level of cathepsin D proteolysis of F42 and F43. Structure-activity relationship studies with analogues of 1a and 1b point to the aldehyde moiety of both compounds being critical to their effects on bMBP structure. The atheronals also cause a reduction in the size of the bMBP-cyt-LUV aggregates, as determined by fluorescence microscopy and dynamic light scattering. These results suggest that formation of an imine between inflammatory-derived aldehydes, which effectively reduces the cationic nature of MBP, can lead to structural changes in MBP and a decrease in myelin stability akin to deimination and as such may make a hitherto unknown contribution to the onset and progression of MS.
authors.affiliation: The Scripps-Oxford Laboratory, Department of Biochemistry, University of Oxford , South Parks Road, Oxford OX1 3QU, UK.
authors.forename: Natalie K
authors.initials: NK
authors.lastname: Cygan
authors.forename: Johanna C
authors.initials: JC
authors.lastname: Scheinost
authors.forename: Terry D
authors.initials: TD
authors.lastname: Butters
authors.forename: Paul
authors.initials: P
authors.lastname: Wentworth
mesh_terms: D000447:Aldehydes / Q000737:chemistry* / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002402:Cathepsin D / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry* / Q000187:drug effects / Q000276:immunology; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000737:chemistry* / Q000378:metabolism*; D055550:Protein Stability / Q000187:drug effects; D017434:Protein Structure, Tertiary; D013499:Surface Properties; D053835:Unilamellar Liposomes / Q000737:chemistry / Q000378:metabolism
chemical_list: D000447:Aldehydes; D016056:Immunodominant Epitopes; D004676:Myelin Basic Protein; D053835:Unilamellar Liposomes; C512174:atheronal-A; C512175:atheronal-B; D002784:Cholesterol; D002402:Cathepsin D