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X-Prolyl dipeptidyl-aminopeptidase activity was found in human urine by a sensitive fluorescence assay in which a new fluorogenic substrate, 7-glycylproline-4-methylcoumarinamide, is used. The Km value was 2.9 X 10(-4) mol/liter, and the optimum pH was 8.7 in glycine-NaOH buffer. The enzyme activity was stable at 4 degrees C for at least five days. On Sephadex G-200 column chromatography, normal human urine showed a main peak with an approximate relative molecular mass of 400 000. The procedure is simple, rapid, and accurate. The enzyme activity in urine of normal adults was: 4.30 +/- 0.13 (SE) (range, approximately 1.84-8.96) micronmol/min per gram of creatinine, and 2.16 +/- 0.09 (SE) (range, approximately 0.38-6.98) micronmol/min per liter of urine.
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Molecular and cellular biochemistry,
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Experientia,
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Chemical & pharmaceutical bulletin,
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Clinical chemistry,
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June 2011,
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