Summary
A new, simple assay to measure peptide hydrolase activity in human small bowel biopsy material is introduced. Amino acids released after peptide hydrolysis are measured enzymatically by means of L-amino acid oxidase. This method has a high sensitivity, is of great simplicity, allows substrates composed of aromatic amino acids to be used, and is suitable for kinetic enzymatic analysis. The spectrum of intestinal peptide hydrolase activities therefore can be greatly amplified. Since not only the supernatent but also the total small bowel biopsy homogenate can be used in the assay to measure peptide hydrolase activity the method opens the possibility to differentiate between brush border and cytoplasmatic peptide hydrolase activity by inhibiting cytoplasmatic peptide hydrolase activity.
Zusammenfassung
Es wird über eine neue, einfache Methode zur Bestimmung der Peptidhydrolasenaktivität in menschlichem Dünndarmbiopsiematerial berichtet. Dabei werden die nach Peptidspaltung freigesetzten Aminosäuren enzymatisch mit L-Aminosäuren-Oxidase bestimmt. Die Methode hat gegenüber früheren Bestimmungsmethoden den Vorteil hoher Empfindlichkeit, ist einfach, erlaubt, Substrate aus aromatischen Aminosäuren zu verwenden und eignet sich für enzymkinetische Untersuchungen. Das Spektrum intestinaler Peptidhydrolasenaktivitäten läßt sich dadurch erheblich erweitern. Da die Bestimmungen der Peptidhydrolasenaktivität im Gesamthomogenat durchgeführt werden können, eröffnet sich die Möglichkeit, durch Hemmung der cytoplasmatischen Peptidhydrolasenaktivität zwischen Bürstensaum- und cytoplasmatischer Aktivität zu unterscheiden.
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Über einen Teil der Ergebnisse wurde auf der 27. Tagung der Deutschen Gesellschaft für Verdauungs- und Stoffwechselerkrankungen, Frankfurt a.M., Oktober 1972, berichtet.
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Caspary, W.F. Intestinale Peptidhydrolasenaktivität in menschlichem Dünndarmbiopsiematerial — eine einfache Bestimmungsmethode. Klin Wochenschr 52, 341–344 (1974). https://doi.org/10.1007/BF01468832
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DOI: https://doi.org/10.1007/BF01468832
Key words
- Peptide hydrolase activity
- Peptidases
- L-Aminoacid-Oxydase
- Intestinal Enzymes
- Brush border enzymes
- Small bowel biopsies