Coenzyme A-independent monoacylglycerol acyltransferase from rat intestinal mucosa.
Rat intestinal mucosa contains high diacylglycerol-synthesizing activity (monoacylglycerol acyltransferase (MGAT) activity) due to monoacylglycerol and fatty acid, independently of coenzyme A and ATP. MGAT activity was purified from rat intestinal mucosa by successive chromatography separations on DEAE-cellulose, CM- Sephadex, and anti-IgG-Sepharose against rat pancreatic lipase. The enzyme was electrophoretically homogeneous, and its molecular weight was 49,000, which is identical with that of rat pancreatic lipase. Immunoblotting analysis with antibody against rat pancreatic lipase showed one immunoreactive protein with an estimated molecular weight of 49,000. The activity of the purified enzyme was completely inhibited by addition of the antibody. Using immunocytochemical techniques, it was found that immunoreactive protein against rat pancreatic lipase was uniformly distributed within the absorptive cells of the intestine but was absent from the microvillar membrane. The MGAT activity of intestinal mucosal homogenate was inhibited by about 65% by addition of antibody against rat pancreatic lipase. Trioleoylglycerol- and dioleoylglycerol-hydrolyzing activities of the purified enzyme and pancreatic lipase were inhibited by addition of intestinal mucosa extract. These results suggest that pancreatic lipase is present in intestinal absorptive cells and that it may contribute to resynthesis of diacylglycerol from monoacylglycerol and fatty acids in these cells.
Tsujita
T
T
Department of Medical Biochemistry, School of Medicine, Ehime University, Ehime, Japan.
Miyazaki
T
T
Tabei
R
R
Okuda
H
H
D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D003065:Coenzyme A / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D005989:Glycerides / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000201:enzymology*; D008049:Lipase / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
D004075:Diglycerides; D005227:Fatty Acids; D005989:Glycerides; D000217:Acyltransferases; C021802:2-acylglycerol O-acyltransferase; D008049:Lipase; D003065:Coenzyme A