Characterization of two dipeptidases purified from hepatic schistosome egg granulomas in mice. Leukotriene D4 hydrolases of granulomatous tissue.

Abstract

Extracts prepared from tissue with granulomatous inflammation experimentally produced in liver of CBA-strain mice showed increased hydrolysis of leukotriene D4 (LTD4), Leu-Leu and Ala-Gly as compared with normal hepatic cells. Two dipeptidases, Leu-Leu dipeptidase and Ala-Gly dipeptidase, were purified from hepatic granulomas, and quantitative conversion of LTD4 into leukotriene E4 (LTE4) by both enzymes was demonstrated. M(r) values of the purified enzymes were 178,000 for Leu-Leu dipeptidase and 183,000 for Ala-Gly dipeptidase. The enzymes showed homogeneity, appearing as a single band on SDS/PAGE, and the M(r) values of the subunits were 56,000 and 57,000 for Leu-Leu and Ala-Gly dipeptidase respectively. The amino acid compositions of the two enzymes differed considerably from each other. The activity of Leu-Leu dipeptidase was inhibited by bestatin and captopril and stabilized with MnCl2. The Km for LTD4 was 25 microM with a V(max.) of 49.0 mumols/min per mg. In contrast, the activity of Ala-Gly dipeptidase was inhibited by cilastatin, cytinylglycine, EDTA and dithiothreitol, and also by captopril. The Km for LTD4 was 5.3 microM with a V(max.) of 50.4 mumols/min per mg. The findings indicate that the conversion of LTD4 into LTE4 by microsomal dipeptidases is elevated during granulomatous tissue reaction. This enzyme activity may become useful for biochemical quantification of the pathological tissue reaction that occurs in organized granulomas.

• Authors: N Sato, Y Ito, T Iida, K Fukuyama, W L Epstein
• Journal: The Biochemical journal (Biochem J) Vol. 284 ( Pt 3) Pg. 885-90 (Jun 15 1992) ISSN: 0264-6021 [Print] England
• PMID: 1622404 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Dipeptides
• Oligopeptides
• Dipeptidases
• leukotriene D4 dipeptidase

Topics

• Amino Acid Sequence
• Animals
• Chromatography, Affinity
• Chromatography, Gel
• Chromatography, High Pressure Liquid
• Chromatography, Ion Exchange
• Dipeptidases (isolation & purification, metabolism)
• Dipeptides (metabolism)
• Electrophoresis, Polyacrylamide Gel
• Female
• Granuloma (enzymology)
• Kinetics
• Liver (enzymology)
• Liver Diseases, Parasitic (enzymology)
• Mice
• Mice, Inbred CBA
• Molecular Sequence Data
• Molecular Weight
• Oligopeptides (metabolism)
• Ovum
• Schistosoma mansoni (physiology)
• Schistosomiasis mansoni (enzymology)
• Substrate Specificity