FIGURE 2.

Pin1 elevates the NF-H phosphorylation by proline directed Ser/Thr kinases. A, purified NF-H was incubated at 30 °C for 2 h with increasing concentrations of GST-Pin1 (lanes 1-5), in the presence of p42/44 MAPK and 1 mm ATP. Reactions were terminated by addition of SDS loading buffer, and proteins were analyzed by 4-12% SDS-PAGE. Blots were probed with monoclonal antibody RT97 that specifically detects p-NF-H. B, densitometry analysis of p-NF-H with data obtained from A. C, reactions were performed as in A with Cdk5/p35 at 30 °C for 2 h. D, densitometry analysis of p-NF-H with data obtained from C. E, reactions were performed as in A with JNK3 at 30 °C for 2 h. F, densitometry analysis of p-NF-H with data obtained from E. Data shown are the mean ± S.E. of four different experiments.