Prorenin activity assay

Human prorenin is activated by controlled digestion until the conversion is complete as evidenced by a shift in molecular weight by SDS PAGE. We do not routinely measure enzymatic activity on each lot but this can be performed using the fluorescent peptide substrate (Arg-Glu(EDANS)-Ile-His-Pro-Phe-His-Leu-Val-Ile-His-Thr-Lys(dabcyl)-Arg) from Molecular Probes (Cat # R-2931). Activity is detected in a microplate format using a SpectroMax Gemini microplate reader. We have done this before and observe a dramatic increase in substarte hydrolysis subsequent to prorenin activation. Prorenin itself has only 1-5% of the enzymatic activity of the renin produced after activation.

Specific activity is determined to be 100 or greater pmoles of substrate hydrolyzed/min/μg.

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