Variable denaturation of ovalbumin by incorporation of amino acid analogs

Denaturation of hen ovalbumin synthesized in a cell-free system was assayed by examining its sensitivity to trypsin. The native ovalbumin resisted digestion by trypsin, and it remained resistant to digestion when some amino acid analogs, including azetidine-2-carboxylic acid and meta-flourotyrosine, were incorporated into its peptide chain. However, when other amino acid analogs such as beta-hydroxyleucine and 4-thiaisoleucine were incorporated during protein systhesis, ovalbumin became very lablie to trypsin. These experiments demonstrate a sensitive system for detecting protein denaturation and suggest a variable effect of different amino acid analogs on the native conformation of a protein.