Soluble neutral maltase--glucoamylase from the small intestine: separation and characterization of components with differing affinity for concanavalin A.
Intestinal maltase with a neutral pH optimum exists in both a brush border membrane-bound form and a soluble form in suckling rat intestine. Previous experiments in our laboratory have shown that the soluble enzyme contains a component which binds much more tightly to concanavalin A (ConA) than solubilized forms of the membrane enzyme. We studied the origin of this component by subjecting neutral, soluble maltase activity to chromatography on Sepharose 4B at age 13, 18 (preweaning), and 25 (postweaning) days. At 13 days, two maltase peaks were obtained with approximate molecular weights of 400 000 (peak I) and 150 000 (peak II). Peak II was less prominent at 18 days and was absent at 25 days. At 13 days, the majority of peak I consisted of material which was bound between 0.025 and 0.05 M alpha-methyl mannoside on gradient elution chromatography of ConA-Sepharose. Peak II contained material which eluted between 0.075 and 0.3 M alpha-methyl mannoside. At 25 days, all of the soluble maltase eluted between 0.025 and 0.04 M alpha-methyl mannoside. Peak I and peak II maltases had similar pH optima and Km's for maltase. Peak II maltase had a fourfold greater activity toward glycogen than peak I maltase with approximately the same activity for palatinose, turanose, and trehalose. Both maltases were precipitated by an antibody raised against adult membrane-bound maltase. Soluble maltase with neutral pH activity in the suckling rat intestine, therefore, consists of two immunologically related isozymes which differ in their molecular weight, their binding by ConA, and their specificity for glycogen. The small isozyme disappears at or about the time of weaning.
D000818:Animals; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000378:metabolism*; D005260:Female; D005087:Glucan 1,4-alpha-Glucosidase / Q000302:isolation & purification* / Q000378:metabolism; D005959:Glucosidases / Q000302:isolation & purification*; D007421:Intestine, Small / Q000201:enzymology*; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity; D014886:Weaning
D007527:Isoenzymes; D003208:Concanavalin A; D005959:Glucosidases; D005087:Glucan 1,4-alpha-Glucosidase