Sugar hydrolases and their arrangement on the rat intestinal microvillus membrane.
The arrangement of the sugar hydrolases, sucrase-isomaltase, maltase, and lactase on the microvillus membrane of rat intestine was investigated by immunological technique. The enzymes were purified essentially free of each other to near homogeneity and antisera of high specificity were obtained against each. Microvillus membranes were prepared routinely in high purity from rat intestine and contained an average 61% protein, 20% lipid, and 19% carbohydrate, with the sugar hydrolases comprising an estimated 20--25% of the membrane protein. The immunoreactivity of membrane-bound sucrase-isomaltase, maltase, and lactase was investigated with antisera demostrating specific reactivity to each, when tested in the presence of other membrane extractives. The membrane-bound enzymes were found in each case to combine with antibody in amounts equivalent to that required to effect precipitation of comparable units of the free enzymes from solution. Preloading membrane vesicles with antibodies to any two of the enzymes did not affect either the immunoreactivity or extractability (by papain or Triton X-100) of the third. The antibody-binding studies indicated an arrangement of these enzymes independent of each other on the membrane surface, in a manner allowing each to maintain a high degree of molecular freedom.
D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000032:analysis* / Q000302:isolation & purification; D005779:Immunodiffusion; D007413:Intestinal Mucosa / Q000201:enzymology*; D007421:Intestine, Small / Q000201:enzymology*; D007700:Kinetics; D008871:Microvilli / Q000201:enzymology* / Q000648:ultrastructure; D051381:Rats; D013394:Sucrase-Isomaltase Complex / Q000032:analysis; D000520:alpha-Glucosidases / Q000032:analysis; D001616:beta-Galactosidase / Q000032:analysis
D006026:Glycoside Hydrolases; D013394:Sucrase-Isomaltase Complex; D000520:alpha-Glucosidases; D001616:beta-Galactosidase