title: Cloning and characterization of Schistosoma mansoni fructose-1,6-bisphosphate aldolase isoenzyme.
abstract: A Schistosoma mansoni cercarial cDNA expression library, constructed in lambda gt11, was screened using the IgG fraction of sera taken from rabbits vaccinated with irradiated cercariae. A positive cDNA clone (1,431 base pairs) was selected and characterized. The amino acid sequence predicted from the cDNA sequence identified a polypeptide of 363 amino acids that showed significant homology to different family members of the enzyme fructose-1,6-bisphosphate aldolase (EC 1.4.2.13). The identity was 66% and 65% with human C and A isoenzymes, respectively. Active sites and substrate-binding determinant analysis suggest that the isolated enzyme in terms of function resembles type A aldolase. The recombinant protein expressed in the vector pGEX-2T was found to be active enzymatically. Antibodies raised against the purified recombinant protein recognized a 40-kDa band in extracts from cercariae, schistosomula (5 and 25 days), adult worms, and eggs. Using immunocytochemistry, aldolase localized to the tegumental region of the adult worms.
authors.affiliation: Theodore Bilharz Research Institute, Cairo, Egypt.
authors.forename: E
authors.initials: E
authors.lastname: El-Dabaa
authors.forename: H
authors.initials: H
authors.lastname: Mei
authors.forename: A
authors.initials: A
authors.lastname: El-Sayed
authors.forename: A M
authors.initials: AM
authors.lastname: Karim
authors.forename: H M
authors.initials: HM
authors.lastname: Eldesoky
authors.forename: F A
authors.initials: FA
authors.lastname: Fahim
authors.forename: P T
authors.initials: PT
authors.lastname: LoVerde
authors.forename: M A
authors.initials: MA
authors.lastname: Saber
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004331:Drosophila melanogaster; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D011817:Rabbits; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D007527:Isoenzymes; D005634:Fructose-Bisphosphate Aldolase