The presence of amyloid beta-protein in the detergent-insoluble membrane compartment of human neuroblastoma cells.
To investigate the intracellular compartmentalization of amyloid beta-protein (Abeta), human neuroblastoma SH-SY5Y cells were fractionated and the Abeta content in each fraction was quantitated by the well-characterized two-site enzyme-linked immunosorbent assay (ELISA). Subcellular fractionation of the cell revealed two distinct pools of Abeta within the cells: a Triton-soluble and a Triton-insoluble pools with the latter being larger than the former. Because Triton insolubility points to caveolae-like domains, we prepared detergent-insoluble, low-density membrane domains from SH-SY5Y cells using two different protocols. The low-density membrane fraction prepared by either protocol was found to contain a substantial proportion of intracellular Abeta40 and Abeta42. These results indicate that the distinct membrane domains are involved in the generation and/or trafficking of Abeta.
Morishima-Kawashima
M
M
Department of Neuropathology, Faculty of Medicine, University of Tokyo, Japan.
Ihara
Y
Y
D016229:Amyloid beta-Peptides / Q000302:isolation & purification* / Q000378:metabolism; D002451:Cell Compartmentation; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003902:Detergents; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D009447:Neuroblastoma / Q000737:chemistry* / Q000378:metabolism; D017830:Octoxynol; D012995:Solubility; D014407:Tumor Cells, Cultured
D016229:Amyloid beta-Peptides; D003902:Detergents; D008565:Membrane Proteins; D017830:Octoxynol