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Clinical Chemistry 24: 1163-1166, 1978;
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Clinical Chemistry, Vol 24, 1163-1166, Copyright © 1978 by American Association for Clinical Chemistry

Excretion of X-prolyl dipeptidyl-aminopeptidase in human urine as determined with a new fluorogenic substrate

T Kato, T Hama, K Kojima, T Nagatsu and S Sakakibara

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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Copyright © 1978 by the American Association for Clinical Chemistry.