Faecal mucinase activity assessed in inflammatory bowel disease using 14C threonine labelled mucin substrate.
Previous studies have shown the presence in faeces of sulphatases, sialidases, glycosidases, and proteases relevant to mucus degradation, but the relative role of these enzymes in the degradation of colonic mucus has been unclear. A total mucinase assay using 14C threonine biologically labelled human colonic mucin as substrate was therefore developed in this study. Faecal mucinase activity of a pooled normal faecal filtrate was capable of removing 80% of the 14C threonine label from mucin within eight hours incubation, but 20% remained intact despite prolonged incubation. The pH profile of mucinase activity is broad (pH 4.5-9.5) suggesting contribution from multiple enzymes. Mucinase activity was reduced by preincubation with 100 micrograms/ml chymostatin (82.8%), 0.5 mg/ml EDTA (91.6%), and 4 g/l bismuth subsalicylate (72.0%). All 55 faecal samples studied contained detectable mucinase activity, measured as dpm release/micrograms protein/hour, which was greater in samples from patients with ulcerative colitis (n = 17, median 52.7, interquartile range 32.9-66.9), than controls (n = 26, 34.4, 26.8-40.4, p < 0.02) or patients with Crohn's disease (n = 12, 35.5, 17.5-55.7, p < 0.05). There was, however, no significant difference in faecal mucinase activity between inactive and active ulcerative colitis. These results suggest that faecal mucinase activity is one factor contributing to the thin mucus layer in ulcerative colitis and represents a potential target for drug treatment.
Dwarakanath
A D
AD
Department of Medicine, University of Liverpool.
Campbell
B J
BJ
Tsai
H H
HH
Sunderland
D
D
Hart
C A
CA
Rhodes
J M
JM
D003093:Colitis, Ulcerative / Q000201:enzymology*; D003424:Crohn Disease / Q000201:enzymology*; D005243:Feces / Q000201:enzymology*; D006801:Humans; D009077:Mucins / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013912:Threonine / Q000378:metabolism
D009077:Mucins; D013912:Threonine; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase