Abstract
The molecular conformation of nuclease P1 in aqueous solution was investigated by measuring the optical rotatory dispersion (ORD) and circular dichroism (CD). The optical rotatory dispersion constant, λc, was 281 nm. The Moffit-Yang parameters, a0 and b0, were -2 and -195, respectively. The ORD spectrum showed a minimum at 234 nm and the reduced mean residue rotation at 233 nm, [m']233, was -5880. The CD spectrum showed a double minimum at 213 and 226 nm and the molecular ellipticity at 222 nm, [θ]222, was -11, 900. From these data, the α-helix content was calculated to be 29 to 31%. The computer fit of CD suggests that the β-structure is about 6% and the random coil is about 63%. The helical structure was found to be quite stable to denaturing reagents such as urea and guanidine hydrochloride. However, removal of zinc atoms from the enzyme resulted in disruption of the helical structure with inactivation.
