A study of the native-denatured (N⇌D) transition in lysozyme I. Detection of the transition by product analyses of protease digests

Abstract

Protease digestion was found to be a convenient probe of the native-denatured conformation transition in lysozyme. Chromatographic analyses of proteolytic digests of lysozyme, Glu-35 ester lysozyme and the complex of lysozyme with the β(1–4)-linked trimer of N-acetylglucosamine indicate that protease digestions proceed via an all-or-none type mechanism and that protease digests only unfolded molecules. The concentration of the unfolded state was reduced by either introduction of one covalent bond in the protein or by association with a substrate analog inhibitor.