Physicochemical and immunological studies of the renatured alpha 1(II) chains and isolated cyanogen bromide peptides of type II collagen.

The alpha 1(II) and cyanogen bromide (CB)1-generated peptides of chick type II collagen were isolated, purified, renatured and examined for their physicochemical and immunological properties. The alpha 1(II) chains and peptides CB-6 through CB-12 (3,000 to 40,000 daltons) formed renatured thermostable products as determined by measurements of reduced viscosity, optical rotation and Stokes radius. Moreover, renatured alpha 1 (II) chains and CB-10 were observed to form segment-long-spacing (SLS) crystallites under appropriate conditions. When examined for immunoreactivity with defined rat polyclonal and mouse monoclonal antibodies to chick type II collagen, conformation-dependent epitopes were detected on renatured alpha 1(II) chains and renatured peptides, CB-8, CB-10 and CB-11. Conformation-independent epitopes were also detected on all CB-peptides in their denatured form. These studies demonstrate that the alpha 1 (II) chains and CB-peptides of chick type II collagen can be efficiently renatured and that the renatured products retain some conformation-dependent epitopes present on the naive molecule.