Brain Levels of Microtubule-Associated Protein τ Are Elevated in Alzheimer's Disease: A Radioimmuno-Slot-Blot Assay for Nanograms of the Protein |
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Authors: | Sabiha Khatoon Inge Grundke-Iqbal Khalid Iqbal |
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Institution: | New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York 10314. |
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Abstract: | The microtubule-associated protein tau, which stimulates the assembly of alpha-beta tubulin heterodimers into microtubules, is abnormally phosphorylated in Alzheimer's disease (AD) brain and is the major component of paired helical filaments. In the present study, the levels of tau and abnormally phosphorylated tau were determined in brain homogenates of AD and age-matched control cases. A radioimmuno-slot-blot assay was developed, using a primary monoclonal antibody, Tau-1, and a secondary antibody, antimouse 125I-immunoglobulin G. To assay the abnormally phosphorylated tau, the blots were treated with alkaline phosphatase before immunolabeling. The levels of total tau were about eightfold higher in AD (7.3 +/- 2.7 ng/micrograms of protein) than in control cases (0.9 +/- 0.2 ng/micrograms), and this increase was in the form of the abnormally phosphorylated protein. These studies indicate that the abnormal phosphorylation--not a decrease in the level of tau--is a likely cause of neurofibrillary degeneration in AD. |
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Keywords: | Radioimmunoslot-blot assay Microtubule-associated protein τ Abnormal phosphorylation Neurofibrillary tangles Paired helical filaments |
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