EXPRESSION OF THE CATALYTIC DOMAIN OF CYCLIC GMP-DEPENDENT PROTEIN-KINASE IN A BACULOVIRUS SYSTEM

The Type I cGMP-dependent protein kinase catalytic domain (residues 336-671 from the I alpha isoform) has been expressed as a cGMP independent kinase in a baculovirus system. Using peptide substrates, the protein retains similar substrate specificity as the native holoenzyme. The recombinant catalytic domain catalyzes the phosphorylation of histone, but does not display the inhibition using non-substrate histones which has been described for the holoenzyme. The catalytic domain is an active kinase in mammalian cells also since vascular smooth muscle cells transfected with the cDNA encoding the catalytic domain display altered morphology. The catalytic domain of G-kinase may be a useful tool for delineating the role of cGMP-mediated protein phosphorylation in cell systems.