Fig. 10.

Autoradiograph showing ³²P-labeled CNBr cleavage fragments of myoIII_Lim. Soluble proteins from lateral optic nerve homogenates that had been phosphorylated with [γ-³²P]ATP in the presence of 8-bromo-cAMP were separated by SDS-PAGE, blotted to PVDF, and stained with Ponceau S. The 122 kDa band was excised and cleaved with CNBr. After the CNBr cleavage fragments were released from the membrane, they were separated on a Tris–Tricine gel and blotted to PVDF. The blots were stained with Coomassie blue R-250 and exposed to autoradiographic film. The two labeled 50 kDa bands shown were cut out and pooled for N-terminal sequencing. The N-terminal sequence was determined as REKFEYL-PL, which matches closely a region in the deduced sequence of myoIII_Lim located near the N terminal of the myosin domain.