Gc (vitamin D-binding protein) binds the 33.5 K tryptic fragment of actin

Limited proteolysis of G-actin was performed with trypsin and chymotrypsin to compare the binding sites for Gc and DNase. DNase I bound to the N-terminal area corresponding to the major cleavage site on G-actin (residues 62–68) and inhibited proteolysis, but did not bind the 33.5K C-terminal fragment (G-actin 33.5) generated. In contrast, Gc did not exert any inhibitory effect upon proteolysis of the intact native G-actin 42.0 molecule, although its presence protected G-actin 33.5 from further proteolysis. This was shown by gel filtration to be due to the formation of complexes between Gc and G-actin 33.5.