Abstract
We report a novel assay method for enterokinase capable of detecting approx. 1 fmol of enzyme. The method depends on quantification of the release of specifically radiolabelled activation peptides from bovine trypsinogen and is unaffected by trypsin inhibitors. The assay is applicable to biological fluids such as serum. The substrate was produced by selective epsilon-amidination of bovine trypsinogen followed by acetylation with [3H]acetic anhydride and deprotection. The assay has been used to study the effects of pH, Ca2+, ionic strength abd glycodeoxycholate on enterokinase activity.
Full text
PDF





Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson L. E., Walsh K. A., Neurath H. Bovine enterokinase. Purification, specificity, and some molecular properties. Biochemistry. 1977 Jul 26;16(15):3354–3360. doi: 10.1021/bi00634a011. [DOI] [PubMed] [Google Scholar]
- Baratti J., Maroux S., Louvard D. Effect of ionic strength and calcium ions on the activation of trypsinogen by enterokinase. A modified test for the quantitative evaluation of this enzyme. Biochim Biophys Acta. 1973 Oct 10;321(2):632–638. doi: 10.1016/0005-2744(73)90206-4. [DOI] [PubMed] [Google Scholar]
- Grant D. A., Hermon-Taylor J. Hydrolysis of artificial substrates by enterokinase and trypsin and the development of a sensitive specific assay for enterokinase in serum. Biochim Biophys Acta. 1979 Mar 16;567(1):207–215. doi: 10.1016/0005-2744(79)90187-6. [DOI] [PubMed] [Google Scholar]
- Grant D. A., Hermon-Taylor J. The purification of human enterokinase by affinity chromatography and immunoadsorption. Some observations on its molecular characteristics and comparisons with the pig enzyme. Biochem J. 1976 May 1;155(2):243–254. doi: 10.1042/bj1550243. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grant D. A., Jones P. A., Magee A. I., Hermon-Taylor J. The biliary excretion of intravenously administered enterokinase [proceedings]. Biochem Soc Trans. 1980 Feb;8(1):55–55. doi: 10.1042/bst0080055. [DOI] [PubMed] [Google Scholar]
- Grant D. A., Magee A. I., Hermon-Taylor J. Optimisation of conditions for the affinity chromatography of human enterokinase on immobilised p-aminobenzamidine. Improvement of the preparative procedure by inclusion of negative affinity chromatography with glycylglycyl-aniline. Eur J Biochem. 1978 Jul 17;88(1):183–189. doi: 10.1111/j.1432-1033.1978.tb12436.x. [DOI] [PubMed] [Google Scholar]
- Grant D. A., Magee A. I., Meeks D., Regan C., Bainbridge D. R., Hermon-Taylor J. Identification of a defence mechanism in vivo against the leakage of enterokinase into the blood. Biochem J. 1979 Dec 15;184(3):619–626. doi: 10.1042/bj1840619. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hadorn B., Hess J., Troesch V., Verhaage W., Götze H., Bender S. W. Role of bile acids in the activation of trypsinogen by enterokinase: disturbance of trypsinogen activation in patients with intrahepatic biliary atresia. Gastroenterology. 1974 Apr;66(4):548–555. [PubMed] [Google Scholar]
- Hanes C. S. Studies on plant amylases: The effect of starch concentration upon the velocity of hydrolysis by the amylase of germinated barley. Biochem J. 1932;26(5):1406–1421. doi: 10.1042/bj0261406. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
- Nureddin A., Inagami T. Chemical modifications of amino groups of trypsin. Biochem Biophys Res Commun. 1969 Sep 10;36(6):999–1005. doi: 10.1016/0006-291x(69)90303-9. [DOI] [PubMed] [Google Scholar]
- Offord R. E. Electrophoretic mobilities of peptides on paper and their use in the determination of amide groups. Nature. 1966 Aug 6;211(5049):591–593. doi: 10.1038/211591a0. [DOI] [PubMed] [Google Scholar]
- Preiser H., Schmitz J., Maestracci D., Crane R. K. Modification of an assay for trypsin and its application for the estimation of enteropeptidase. Clin Chim Acta. 1975 Mar 10;59(2):169–175. doi: 10.1016/0009-8981(75)90025-x. [DOI] [PubMed] [Google Scholar]
- Rinderknecht H., Engeling E. R., Bunnell M. J., Geokas M. C. A sensitive assay for human enterokinase and some properties of the enzyme. Clin Chim Acta. 1974 Jul 31;54(2):145–160. doi: 10.1016/0009-8981(74)90232-0. [DOI] [PubMed] [Google Scholar]
- Rinderknecht H., Friedman R. Effect of bile acids and ionic strength on trypsinogen activation by human enteropeptidase. Biochim Biophys Acta. 1978 Jul 7;525(1):200–208. doi: 10.1016/0005-2744(78)90215-2. [DOI] [PubMed] [Google Scholar]
- WALSH K. A., NEURATH H. TRYPSINOGEN AND CHYMOTRYPSINOGEN AS HOMOLOGOUS PROTEINS. Proc Natl Acad Sci U S A. 1964 Oct;52:884–889. doi: 10.1073/pnas.52.4.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
