title: Comparison of sucrase-free isomaltase with sucrase-isomaltase purified from the house musk shrew Suncus murinus.
abstract: We purified sucrase-isomaltase and sucrase-free isomaltase from a normal and a sucrase-deficient line, respectively, of the house musk shrew Suncus murinus and examined the effects of mutation on enzyme structure and activities. Recent cDNA cloning studies have predicted that sucrase-free mutant isomaltase lacks the C-terminal 69 amino acids of normal isomaltase, as well as the entire sucrase. On SDS-polyacrylamide gel electrophoresis purified sucrase-free isomaltase gave a single protein band of 103 kDa, while sucrase-isomaltase gave two major protein bands of 106 and 115 kDa. The 115, but not 106, kDa band was quite similar to the 103 kDa band on Western blotting with Aleuria aurantia lectin and antibody against shrew sucrase-isomaltase, suggesting that the 115 and 103 kDa bands are due to normal and mutant isomaltases, respectively, in accordance with the above prediction. Purified isomaltase and sucrase-isomaltase were similar in Km and Vmax (based on isomaltase mass) values for isomaltose hydrolysis and in inhibition of isomaltase activity by antibody against rabbit sucrase-isomaltase, suggesting that the enzymatic properties of isomaltase are mostly unaffected by mutation.
authors.affiliation: Department of Biomolecular Science, Faculty of Engineering, Gifu Univeristy, Japan. takesue@cc.gifu-u.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Takesue
authors.forename: K
authors.initials: K
authors.lastname: Yokota
authors.forename: S
authors.initials: S
authors.lastname: Oda
authors.forename: S
authors.initials: S
authors.lastname: Takesue
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D005260:Female; D008297:Male; D009834:Oligo-1,6-Glucosidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012788:Shrews; D013379:Substrate Specificity; D013393:Sucrase / Q000378:metabolism*; D013394:Sucrase-Isomaltase Complex / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D013394:Sucrase-Isomaltase Complex; D009834:Oligo-1,6-Glucosidase; D013393:Sucrase