PINCH in the Cellular Stress Response to Tau-Hyperphosphorylation
Figure 4
hp-Tau interacts with the first and second LIM domains of PINCH.
A) LIMS-specific deletion mutations predict that LIM1 and 2 domains of PINCH1 bind to hp-Tau. Expression plasmids for mutants (ΔLIMS 1–5) and full-length PINCH each with a FLAG-tag were generated via a PCR based strategy. Neuronal lysates were immunoprecipitated with anti-FLAG antibody and reacted with A) anti-hp-Tau (AT8), B) Nck-2 or E) MAP2 antibodies. Arrows indicate immunoreactive bands. FL, full length; ΔLIM, delta LIM indicates the LIM domain that was deleted; FLAG, epitope N-DYKDDDDK-C tag; Nck2, cytoplasmic adaptor protein that interacts with PINCH LIM4; MAP2, microtubule associated protein-2.