Vogelsberg-Ragaglia V, Bruce J, Richter-Landsberg C, Zhang B, Hong M, Trojanowski JQ, Lee VM.
Distinct FTDP-17 missense mutations in tau produce tau aggregates and other pathological phenotypes in transfected CHO cells.Mol Biol Cell.
REAGENTS/MATERIAL: Site-directed mutagenesis (Quikchange kit; Stratagene) was used to created N279K, delta280K, P301L, S305N, V337M, R406W and triple mutation, VPR containing P301L, V337M and R406W) in Tau40 isoform. pSG5 vector (Stratagene) and Pfu DNA polymerase were used
Rabbit polyclonal anti-tau 17026 (Pharmacia Biotech) for immunoprecipitation, 1:500 dilution.
Alpha-tubulin MAb (Blose et al) for Immunofluorescence
Fluroescent-labeled donkey anti-rabbit and Texas Red-labeled donkey anti-mouse IgG (Jackson Laboratories)
FUTURE DIRECTION: Whether reduced binding of tau to MTs is an initiating even that leads to the formation of abnormal tau filaments Further experiments using CHO cell mutant tau model system to study the mechanisms leading to the formation of tau pathology in AD.