FIGURE 4.

In vitro phosphorylation of CRMP2wt-(1–69) and CRMP2Y32F-(1–69). A, alignment of amino acid sequences of rat CRMP1–5. Sequence of rat CRMP1–5, amino acids 25–50. Black line indicates that the epitope of the functional blocking antibody (1) Tyr³² is included in this region. B, in vitro phosphorylation of CRMP2wt-(1–69) or the CRMP2Y32F-(1–69) mutant. In vitro kinase assay was performed using purified Fyn and CRMP2wt-(1–69) or CRMP2Y32F-(1–69) with or without ATP (0.5 mm). Each reaction mixture was subjected to SDS-PAGE and immunoblot analysis with anti-phosphotyrosine antibody (4G10). The same samples were separated and stained by Coomassie Brilliant Blue (CBB) to show that relatively similar amounts of CRMP2 were loaded.