Cloning and characterization of the Schistosoma japonicum aspartic proteinase involved in hemoglobin degradation.
A cDNA encoding a Schistosoma japonicum aspartic proteinase was cloned, sequenced, and found to encode a zymogen of 380 amino acid residues, and its gene was shown to be present as a single copy in the S. japonicum genome. Identity comparisons showed that the enzyme (Sjpasp) was most closely related to the cathepsin Ds. The deduced amino acid sequence has four potential glycosylation sites, two of which are in identical positions to the two glycosylation sites of human kidney lysosomal cathepsin D. Furthermore, all four disulfide bonds found in mammalian cathepsin D sequences are present in Sjpasp, although the beta-hairpin (loop 3), which is cleaved during maturation of vertebrate cathepsin Ds to yield light and heavy chain subunits, is absent from Sjpasp. While most residues involved in substrate specificity and catalysis of aspartic proteinases are preserved in Sjpasp, several residues in these regions exhibit changes that may result in a novel substrate specificity. Aspartic proteinase activity is present in extracts of adult S. japonicum and Schistosoma mansoni and in culture media in which schistosomes were maintained and was capable of digesting hemoglobin. The schistosome aspartic proteinase may play a pivotal role in the catabolism of hemoglobin obtained from host erythrocytes.
Becker
M M
MM
Molecular Parasitology Unit, Queensland Institute of Medical Research, Royal Brisbane Hospital, Australia.
Harrop
S A
SA
Dalton
J P
JP
Kalinna
B H
BH
McManus
D P
DP
Brindley
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015139:Blotting, Southern; D002402:Cathepsin D / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018509:DNA, Helminth / Q000737:chemistry; D017238:Genes, Helminth; D006031:Glycosylation; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012549:Schistosoma japonicum / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D012550:Schistosoma mansoni / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014714:Vertebrates
D018076:DNA, Complementary; D018509:DNA, Helminth; D006454:Hemoglobins; D016282:Aspartic Acid Endopeptidases; D002402:Cathepsin D