Phosphorylation of calponin by PKC is blocked by F-actin in vitro.

ABSTRACT

Polyclonal antibody against phosphorylated calponin was raised in rabbits by application of the peptide corresponding to residues 183-195 of calponin phosphorylated by protein kinase C. When calponin was incubated with protein kinase C, only free calponin was recognized by this antibody and calponin of native thin filament or that binding to F-actin did not. In experiments done using [gamma-32P] ATP, no radioactivity was detected except for free calponin. Calponin phosphorylation was suppressed in an actin dose-dependent manner and the phosphorylation of calponin was completely blocked when the actin molar ratio to calponin exceeded 10. These data suggest that phosphorylation of calponin by protein kinase C was apparently blocked by F-actin.