Conjugated bile salts regulate turnover of rat intestinal brush border membrane hydrolases.
The mechanisms whereby the conjugated bile salts regulate the activities of the brush border membrane hydrolases and its physiological significance were investigated in rat small intestine, and comparisons were made with the action of pancreatic protease. Rat brush border membrane proteins were metabolically labeled with [35S]methionine, and isolated brush border membrane was incubated with taurocholate or pancreatic elastase. The activity of solubilized hydrolases was assayed and the molecular forms of the hydrolases were examined by SDS-PAGE. The activity and protein bands of alkaline phosphatase and sucrase-isomaltase were solubilized by taurocholate, while alkaline phosphatase was not solubilized by elastase. Solubilized sucrase-isomaltase molecules were proteolytically degraded by elastase, whereas the intact molecule of sucrase-isomaltase was solubilized by taurocholate. Next the physiological role of bile salts in brush border membrane hydrolase turnover were investigated using metabolic labeling of brush border membrane hydrolase and immunoprecipitation in biliary diversion rats. After three days of biliary diversion, a significant increase in alkaline phosphatase activity was observed. Although synthesis of alkaline phosphatase in biliary diversion rats was similar to that observed in control rats, biliary diversion rats showed 1.5-fold slower turnover of alkaline phosphatase when compared with control rats. These results suggest that conjugated bile salts in the intestinal lumen may cause a rapid turnover of brush border membrane hydrolases, which may be increased by the enhanced enzyme degradation. The mechanisms for the enhanced degradation appeared to be solubilization of hydrolases caused by the detergent activity of bile salts. Therefore, conjugated bile salts may play an important physiological role in the regulation of expression of the protease-resistant enzymes such as alkaline phosphatase.
Shiozaki
H
H
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Yoshioka
M
M
Miura
S
S
Imaeda
H
H
Morita
A
A
Asakura
H
H
Tsuchiya
M
M
Ishii
H
H
D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism; D000626:Aminopeptidases / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006867:Hydrolases / Q000032:analysis / Q000378:metabolism*; D066298:In Vitro Techniques; D007583:Jejunum / Q000201:enzymology* / Q000648:ultrastructure; D008297:Male; D008871:Microvilli / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar; D012995:Solubility; D013394:Sucrase-Isomaltase Complex / Q000032:analysis / Q000378:metabolism; D013462:Sulfur Radioisotopes
D001647:Bile Acids and Salts; D013462:Sulfur Radioisotopes; D006867:Hydrolases; D000469:Alkaline Phosphatase; D013394:Sucrase-Isomaltase Complex; D000626:Aminopeptidases