title: Fine mapping of the amyloid β-protein binding site on myelin basic protein.
abstract: The assembly and deposition of amyloid β-protein (Aβ) in brain is a key pathological feature of Alzheimer's disease and related disorders. Factors have been identified that can either promote or inhibit Aβ assembly in brain. We previously reported that myelin basic protein (MBP) is a potent inhibitor of Aβ fibrillar assembly [Hoos, M. D., et al. (2007) J. Biol. Chem. 282, 9952-9961; Hoos, M. D., et al. (2009) Biochemistry 48, 4720-4727]. Moreover, the region on MBP responsible for this activity was localized to the 64 N-terminal amino acids (MBP1-64) [Liao, M. C., et al. (2010) J. Biol. Chem. 285, 35590-35598]. In the study presented here, we sought to further define the site on MBP1-64 involved in this activity. Deletion mapping studies showed that the C-terminal region (residues 54-64) is required for the ability of MBP1-64 to bind Aβ and inhibit fibril assembly. Alanine scanning mutagenesis revealed that amino acids K54, R55, G56, and K59 within MBP1-64 are important for both Aβ binding and inhibition of fibril assembly as assessed by solid phase binding, thioflavin T binding and fluorescence, and transmission electron microscopy studies. Strong spectral shifts are observed by solution nuclear magnetic resonance spectroscopy of specific N-terminal residues (E3, R5, D7, E11, and Q15) of Aβ42 upon the interaction with MBP1-64. Although the C-terminal region of MBP1-64 is required for interactions with Aβ, a synthetic MBP50-64 peptide was itself devoid of activity. These studies identify key residues in MBP and Aβ involved in their interactions and provide structural insight into how MBP regulates Aβ fibrillar assembly.
authors.affiliation: Departments of Neurosurgery, Stony Brook University, Stony Brook, New York 11794, USA.
authors.forename: Annmarie E
authors.initials: AE
authors.lastname: Kotarba
authors.forename: Darryl
authors.initials: D
authors.lastname: Aucoin
authors.forename: Michael D
authors.initials: MD
authors.lastname: Hoos
authors.forename: Steven O
authors.initials: SO
authors.lastname: Smith
authors.forename: William E
authors.initials: WE
authors.lastname: Van Nostrand
mesh_terms: D000409:Alanine / Q000235:genetics / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D052160:Benzothiazoles; D001665:Binding Sites; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D046529:Microscopy, Electron, Transmission; D016297:Mutagenesis, Site-Directed; D004676:Myelin Basic Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013844:Thiazoles
chemical_list: D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; C499048:MBP protein, human; D004676:Myelin Basic Protein; D010446:Peptide Fragments; D013844:Thiazoles; C009462:thioflavin T; D000409:Alanine