Binding of the Alzheimer amyloid beta-peptide to neuronal cell membranes by fluorescence correlation spectroscopy.
The deposition of the Alzheimer amyloid beta-peptide (Abeta) fibrils in brain is a key step in Alzheimer's disease. The aggregated Abeta is found to be toxic to neurons since cells die when the aggregated Abeta is added to the cell culture medium. However, target of action of Abeta to cells is unknown. We have applied the fluorescence correlation spectroscopy (FCS) technique to study the existence of a receptor or target molecule for the Alzheimer amyloid beta-peptide (Abeta) in cultured human cerebral cortical neurons. FCS measurement of the fluorophore rhodamine-labeled Abeta (Rh-Abeta) shows diffusion times: 0.1 ms, 1.1 ms and 5.9 ms. Thus, 0.1 ms corresponds to the unbound Rh-Abeta, and 1.1 ms and 5.9 ms correspond to slowly diffusing complexes of Rh-Abeta bound to a kind of receptor or target molecule for Abeta. Addition of excess non-labeled Abeta is accompanied by a competitive displacement, showing that the Abeta binding is specific. Full saturation of the Abeta binding is obtained at nanomolar concentrations, indicating that the Abeta binding is of high affinity. The notion that using FCS we have found a kind of receptor or target molecule for Abeta makes an important point that Abeta kills cells possibly by affecting cell membranes via a receptor or target molecule. This study is of highly significance since it suggests that Abeta possibly affects neuronal cell membranes of Alzheimer patients via a receptor or target molecule.
Hossain
Shakil
S
Department of Medical Biochemistry and Biophysics, Scheeles väg 2, Karolinska Institute, S-171 77 Stockholm, Sweden.
Grande
Mirtha
M
Ahmadkhanov
Galib
G
Pramanik
Aladdin
A
D016229:Amyloid beta-Peptides / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002525:Cerebellar Cortex / Q000166:cytology; D005456:Fluorescent Dyes; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D011485:Protein Binding; D012235:Rhodamines; D013050:Spectrometry, Fluorescence / Q000379:methods
D016229:Amyloid beta-Peptides; D005456:Fluorescent Dyes; D008565:Membrane Proteins; D010446:Peptide Fragments; D012235:Rhodamines; C078592:amyloid beta-protein (1-40)