title: K3 fragment of amyloidogenic beta(2)-microglobulin forms ion channels: implication for dialysis related amyloidosis.
abstract: Beta(2)-microglobulin (beta(2)m) amyloid deposits are linked to dialysis-related amyloidosis (DRA) in hemodialysis patients. The mechanism by which beta(2)m causes DRA is not understood. It is also unclear whether only the full-length beta(2)m induces pathophysiology or if proteolytic fragments are sufficient for inducing this effect. Ser20-Lys41 (K3) is a digestion fragment of full-length beta(2)m. Solid state NMR (ssNMR) combined with X-ray diffraction and atomic force microscopy (AFM) revealed the characteristic oligomeric amyloid conformation of the U-turn beta-strand-turn-beta-strand motif stacked in parallel and stabilized by intermolecular interactions also shown by Abeta(9-40)/Abeta(17-42) and the CA150 WW domain. Here we use the K3 U-turn atomic coordinates and molecular dynamic (MD) simulations to model K3 channels in the membrane. Consistent with previous AFM imaging of other amyloids that show channel-like structures in the membrane, in the simulations K3 also forms ion channels with 3-6 loosely attached mobile subunits. We carry out AFM, single channel electrical recording, and fluorescence imaging experiments. AFM images display 3D ion channel topography with shapes, morphologies, and dimensions consistent with the theoretical model. Electrical conductance measurements indicate multiple single channel conductances, suggesting that various K3 oligomer sizes can constitute the channel structure. Fluorescence measurements in kidney cells show channel-mediated cell calcium uptake. These results suggest that the beta(2)m-induced DRA can be mediated by ion channels formed by its K3 fragment. Because the beta-strand-turn-beta-strand motif appears to be a universal amyloid feature, its ability to form ion channels further suggests that the motif may play a generic role in toxicity.
authors.affiliation: Center for Nanomedicine and Department of Medicine, University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Mirela
authors.initials: M
authors.lastname: Mustata
authors.forename: Ricardo
authors.initials: R
authors.lastname: Capone
authors.forename: Hyunbum
authors.initials: H
authors.lastname: Jang
authors.forename: Fernando Teran
authors.initials: FT
authors.lastname: Arce
authors.forename: Srinivasan
authors.initials: S
authors.lastname: Ramachandran
authors.forename: Ratnesh
authors.initials: R
authors.lastname: Lal
authors.forename: Ruth
authors.initials: R
authors.lastname: Nussinov
mesh_terms: D000682:Amyloid / Q000378:metabolism*; D000686:Amyloidosis / Q000209:etiology* / Q000378:metabolism*; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004553:Electric Conductivity; D006801:Humans; D007473:Ion Channels / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism; D018625:Microscopy, Atomic Force; D056004:Molecular Dynamics Simulation; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D006435:Renal Dialysis / Q000009:adverse effects*; D001613:beta 2-Microglobulin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000682:Amyloid; D007473:Ion Channels; D008051:Lipid Bilayers; D010446:Peptide Fragments; D001613:beta 2-Microglobulin; D002118:Calcium