Cloning and characterization of Schistosoma mansoni fructose-1,6-bisphosphate aldolase isoenzyme.
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.
El-Dabaa
E
E
Theodore Bilharz Research Institute, Cairo, Egypt.
Mei
H
H
El-Sayed
A
A
Karim
A M
AM
Eldesoky
H M
HM
Fahim
F A
FA
LoVerde
P T
PT
Saber
M A
MA
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
D018076:DNA, Complementary; D018509:DNA, Helminth; D007527:Isoenzymes; D005634:Fructose-Bisphosphate Aldolase