Alonso A, Zaidi T, Novak M, Grundke-Iqbal I, Iqbal K. Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments/straight filaments. Proc Natl Acad Sci U S A. 2001 Jun 5;98(12):6923-8. PubMed Abstract

REAGENTS/MATERIAL:
Constructs encoding different human brain t isoforms were subcloned, and recombinant proteins were purified as described in Singh TJ et al. Constructs of t fragments were generated, expressed in Escherichia coli, and purified as described in Sanger et al. and Novak et al. AD P-t and normal t were purified from frozen human brains as described in Kopke E et al. AD P-t was deglycosylated within 4 units/ml endoglycosidase F/N-glycosidase F (Boehringer Mannheim) as described in Wang et al. The dephosphorylation of AD P-t with AP was carried out as described in Alonso AC et al. Deglycosylation was detected by Western blots with lectin Galanthus nivalas agglutinin (GNA, detects terminally linked mannose) and peanut agglutinin (PNA, detects galactose-b(1-3)-N-acetylgalactosamine

FUTURE DIRECTION:
Further studies needed on the mechanisms of abnormal hyperphosphorylation of ts in AD, FTDP-17, and related tauopathies, as well as the inhibition of these processes.