Modified proenzymes as substrates for proteolytic enzymes
AssigneeNederlandse Organisatie voor toegepast-natuurwetenschappelijk onderzoek TNO
InventorJohan Hendrikus Verheijen
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Abstract
Detection or determination of a protease in a sample by incubating the sample with a substrate of the protease and observing proteolytic cleavage of said substrate. The substrate is a modified proenzyme containing a recognition site, e.g., an activation site, cleavable by said protease. Proteolytic cleavage of the modified proenzyme is detected by observing the resulting activity using a suitable substrate of the activated proenzyme. The protease may be e.g. an aspartic protease or a metalloprotease, and the modified proenzyme e.g. pro-urokinase having a mutant activation site which is cleavable by the protease to be determined.
Claims30
A method of determining a protease, or its precursor after activation, in a sample, comprising incubating the sample with a substrate of said protease, determining proteolytic cleavage of said substrate, and correlating data obtained therefrom in order to determine the protease, wherein said substrate is a modified proenzyme containing a recognition site which is cleavable by said protease.
A modified proenzyme derived from a proenzyme by providing it with a recognition site which is cleavable by a protease different from the one which activates the unmodified proenzyme.
Dependencies are inferred from the claim text.