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Specificity of urinary kallikrein activity determination with an amidolytic assay


, : Specificity of urinary kallikrein activity determination with an amidolytic assay. Revista Espanola de Fisiologia 39(3): 225-230

The specificity of a method for the determination of the urinary kallikrein activity based on the amidolytic hydrolysis of the synthetic substrate H-D-valyl-leucyl-arginine-p-nitroanilide has been studied. Results show a good specificity for kallikrein. The amidolytic activity was totally inhibited by aprotinin. The urinary amidolytic activity showed identical dependence on changes in the ionic strength and pH of the incubation medium as the highly purified kallikrein. The sensitivity of the assay was 0.6 nkat/l. The intra-assay coefficient of variation was 4% and the inter-assay coefficient of variation was 10%. The percentage of recovery was between 100 and 110% indicating that no activators or inhibitors of the kallikrein activity on this substrate were present in the urinary samples. Values obtained in presumably healthy subjects were 0.14 .+-. 0.08 pkat/s in man (range: 0.05-0.27 pkat/s) and 0.49 .+-. 0.11 pkat/s in rat (range: 0.35-0.58 pkat/s).

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Related references

Camps, J.; Costa, J.; Pérez-Ayuso, R.M.; Rimola, A.; Arroyo, V.; Gaya, J., 1983: Specificity of the determination of urinary kallikrein activity using an amidolytic assay. The specificity of a method for the determination of the urinary kallikrein activity based on the amidolytic hydrolysis of the synthetic substrate H-D-valyl-leucyl-arginine-p-nitroanilide has been studied. Results show a good specificity for kalli...

Briseid, K.; Hoem, N.O.e; Johannesen, S.; Haug, K., 1995: Amidolytic assay of factor XI in human plasma-significance of kallikrein for the activity measured. Factor XI (FXI) deficiency is associated with an abnormal bleeding state. The extent of bleeding does not correlate well with the plasma concentration of FXI, and it has been suggested that also unknown factors interfere with the bleeding tendency...

Briseid, K.; Hoem, N.O.; Johannesen, S.; Haug, K., 1995: Amidolytic assay of factor XI in human plasma--significance of kallikrein for the activity measured. Factor XI (FXI) deficiency is associated with an abnormal bleeding state. The extent of bleeding does not correlate well with the plasma concentration of FXI, and it has been suggested that also unknown factors interfere with the bleeding tendency...

Bönner, G.; Marin-Grez, M., 1981: Measurement of kallikrein activity in urine of rats and man using a chromogenic tripeptide substrate. Validation of the amidolytic assay by means of bradykinin radioimmunoassay. Urinary kallikrein activity in rats and man was measured by an amidolytic assay using the chromogenic tripeptide D-valyl-leucyl-arginine-p-nitroanilide (S 2266). The sensitivity of the assay permits the measurement of 3.5 to 250 mKU of purified po...

Colman R.W., 1982: Assay of pre kallikrein in human plasma lack of correlation of coagulant assay with esterolytic amidolytic and immunochemical assays. Clinical Research: 561a

Liu C.Y.; Colman R.W., 1976: Human plasma pre kallikrein new method for purification and application of a sensitive amidolytic assay. Circulation Supplement 54(2): II-200

Berthoud V.; Corthorn J., 1985: Specificity of the amidase method in the determination of urinary kallikrein of the rat. Archivos de Biologia y Medicina Experimentales 18(2): R117

Berthoud, V.M.; Corthorn, J.L., 1987: Specificity of the amidase and kininogenase methods for the determination of rat urinary kallikrein. The specificity of the amidase and kininogenase methods for determining rat urinary kallikrein was studied. Male and female rat urine was employed. Esterase A1, A2 and kallikrein were separated by DEAE-Sephadex A-50 chromatography. Esterase A1 sho...

Girolami J.P.; Pecher C.; Bascands J.L.; Cabos G.; Adam A.; Suc J.M., 1989: Purification of human active urinary kallikrein comparative inhibition studies of kininogenase and amidolytic activities. Large scale purification of human active urinary kallikrein is described. The final preparation was found homogeneous by means of SDS Page electrophoresis, amino acid composition and N-terminal analysis. The apparent molecular weight, determined o...

Meani, A.; Fabris, C.; Vianello, D.; Piccoli, A.; Del Favero, G.; Farini, R., 1985: A kinetic assay for human urinary kallikrein determination. A simple kinetic method for human urinary killikrein determination is proposed. In this assay, the release of p-nitroaniline from the chromogenic substrate S-2266 [H-D-Val-Leu-Arg-pNA] at 37.degree. C and pH 8.2 is followed spectrophotometrically...