title: Brush-border disaccharidase synthesis in infant pigs measured in vivo with [2H3]leucine.
abstract: Conscious unrestrained piglets were fasted overnight and infused intravenously with [2H3]leucine for 6 h. Sucrase isomaltase and lactase phlorizin hydrolase were immunoprecipitated from jejunal mucosal membranes, and the immunoprecipitates were electrophoresed on polyacrylamide gels. Bands corresponding to the pro and mature isoforms of both enzymes were acid hydrolyzed. [2H3]leucine isotopic enrichment was measured by gas chromatography-mass spectrometry using negative chemical ionization. Plasma leucine reached isotopic steady state within 90 min. The isotopic enrichment of mucosal leucine was 73% of that of plasma leucine. The high mannose and complex glycosylated forms of prolactase were in isotopic equilibrium, and their isotopic enrichment was 94% of mucosal leucine. The fractional synthesis rates of total and membrane protein were 0.45 and 0.65 days-1, whereas the processing rates of mature lactase, sucrase, and isomaltase were 0.90, 0.23, and 0.21 days-1, respectively. Approximately 65% of the label in the sucrase isomaltase immunoprecipitate was in the complex glycosylated precursor, whereas 73% of the label in lactase phlorizin hydrolase was in the mature (160 kDa) form. We conclude that the low rate of brush-border sucrase synthesis reflects a slow rate at which the complex glycosylated precursor is processed to the brush-border form.
authors: Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030.
authors: M A
authors: MA
authors: Dudley
authors: F
authors: F
authors: Jahoor
authors: D G
authors: DG
authors: Burrin
authors: P J
authors: PJ
authors: Reeds
mesh_terms: D000818:Animals; D007583:Jejunum / Q000378:metabolism* / Q000648:ultrastructure; D010694:Lactase-Phlorizin Hydrolase / Q000096:biosynthesis*; D007930:Leucine / Q000378:metabolism*; D008297:Male; D008871:Microvilli / Q000378:metabolism; D013394:Sucrase-Isomaltase Complex / Q000096:biosynthesis*; D013552:Swine
chemical_list: D013394:Sucrase-Isomaltase Complex; D010694:Lactase-Phlorizin Hydrolase; D007930:Leucine