title: Kinetics of amyloid beta monomer-to-oligomer exchange by NMR relaxation.
abstract: Recent studies have implicated non-fibrillar oligomers of the amyloid beta (Abeta) peptide as the primary toxic species in Alzheimer's disease. Detailed structural and kinetic characterization of these states, however, has been difficult. Here we use NMR relaxation measurements to address the kinetics of exchange between monomeric and large, polymorphic oligomeric species of Abeta(1-40). (15)N and (1)H(N) R(2) data at multiple magnetic fields were recorded for several peptide concentrations subsequent to the establishment of a stable pseudo-equilibrium between monomeric and NMR-invisible soluble oligomeric species. The increase in (15)N and (1)H(N) R(2) rates as a function of protein concentration is independent of nucleus and magnetic field and shows only a small degree of variation along the peptide chain. This phenomenon is due to a lifetime broadening effect arising from the unidirectional conversion of monomer to the NMR-invisible oligomeric species ("dark" state). At a total Abeta(1-40) concentration of 300 microM, the apparent first-order rate constant for this process is approximately 3 s(-1). Fitting the McConnell equations for two dipolar-coupled spins in two-site exchange to transfer-of-saturation profiles at two radiofrequency field strengths gives an estimate for k(off) of 73 s(-1) and transiently bound monomer (1)H(N) R(2) rates of up to 42,000 s(-1) in the tightly bound central hydrophobic region and approximately 300 s(-1) in the disordered regions, such as the first nine residues. The fraction of peptide within the "dark" oligomeric state undergoing exchange with free monomer is calculated to be approximately 3%.
authors.affiliation: Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0520, USA.
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Fawzi
authors.forename: Jinfa
authors.initials: J
authors.lastname: Ying
authors.forename: Dennis A
authors.initials: DA
authors.lastname: Torchia
authors.forename: G Marius
authors.initials: GM
authors.lastname: Clore
mesh_terms: D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000378:metabolism*; D002021:Buffers; D007700:Kinetics; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D012996:Solutions
chemical_list: D016229:Amyloid beta-Peptides; D002021:Buffers; D010446:Peptide Fragments; D012996:Solutions; C078592:amyloid beta-protein (1-40)