Microtubule assembly competence analysis of freshly-biopsied human tau, dephosphorylated tau, and Alzheimer tau

T. D. Garver, R. A.W. Lehman, Melvin Billingsley

Research output: Contribution to journalArticle

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Abstract

Phosphorylation of the microtubule-associated protein tau regulates its binding to microtubules; highly phosphorylated tau is also a prime component of paired helical filaments (PHFs) of Alzheimer's disease (AD). Tau from freshly biopsied human, monkey, and rat brain share similar electrophoretic mobility patterns and overlapping phosphorylated epitopes when compared to AD tau isolated from AD brain. We compared the microtubule reassembly competence of fresh isolates of phosphorylated tau to that of maximally dephosphorylated tau and tau from AD brain. A rapid procedure was developed which permitted the enrichment of phosphorylated and dephosphorylated tau from human biopsies in the absence of protein kinase and phosphatase activity. Microtubule assembly assays, using a spectrophotometric measure and purified bovine brain tubulin, were used to correlate assembly competence with states of tau electrophoretic mobility. Maximally dephosphorylated human biopsy-derived tau and monkey tau were assembly competent; tau from AD brain was virtually unable to direct microtubule assembly. Unmodified, biopsy-derived tau from non-AD brain was intermediate in assembly competence relative to AD tau and dephosphorylated tau. Several lines of evidence were used to correlate phosphorylation states of tau with microtubule assembly. First, in vitro dephosphorylation of human biopsy-derived tau with either PP2A or PP2B alone or in combination led to increasing assembly competence as the electrophoretic mobility of tau increased. Second, addition of the protein phosphatase inhibitor okadaic acid (10 μM) to brain-slice preparations slowed electrophoretic mobility of tau and decreased binding competence. We suggest that tau derived from freshly-biopsied brain exists in a range of phosphorylated states, and that dephosphorylation by PP2A and/or PP2B increases microtubule assembly competence.

Original languageEnglish (US)
Pages (from-to)12-20
Number of pages9
JournalJournal of Neuroscience Research
Volume44
Issue number1
DOIs
StatePublished - Apr 22 1996

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Microtubules
Mental Competency
Alzheimer Disease
Brain
Biopsy
Phosphoprotein Phosphatases
Haplorhini
Phosphorylation
Okadaic Acid
Microtubule-Associated Proteins
Brain Diseases
Tubulin
Protein Kinases
Epitopes

All Science Journal Classification (ASJC) codes

  • Cellular and Molecular Neuroscience

Cite this

Garver, T. D. ; Lehman, R. A.W. ; Billingsley, Melvin. / Microtubule assembly competence analysis of freshly-biopsied human tau, dephosphorylated tau, and Alzheimer tau. In: Journal of Neuroscience Research. 1996 ; Vol. 44, No. 1. pp. 12-20.
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Microtubule assembly competence analysis of freshly-biopsied human tau, dephosphorylated tau, and Alzheimer tau. / Garver, T. D.; Lehman, R. A.W.; Billingsley, Melvin.

In: Journal of Neuroscience Research, Vol. 44, No. 1, 22.04.1996, p. 12-20.

Research output: Contribution to journalArticle

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