The correlation of ribonuclease activity with specific aspects of tertiary structure.

Various physical parameters of bovine pancreatic ribonuclease and of some of its derivatives prepared by oxidation or reduction of disulfide bridges, methylation, and proteolytic digestion, have been investigated. Ultraviolet spectrophotometric measurements appear to establish a correlation between specific spectral properties of the materials and their enzymic activity. Viscometric and optical rotary studies, on the other hand, indicate that minor modifications in secondary structure may occur without detectable inactivation. Polyvalent anions and polyanions almost completely prevent the unfolding and spectral shift effect of 8 M urea, and of guanidium ions at concentrations between 1 and 3 M. These findings suggest that the full activity of ribonuclease, in the presence of such denaturing agents, is due to a refolding of the protein under the influence of ribonucleic acid.

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