Cloning and molecular characterization of calpain, a calcium-activated neutral proteinase, from different strains of Schistosoma japonicum.
cDNA coding for calpain of Schistosoma japonicum were cloned and sequenced, and serological basis of host responses to calpain were analyzed. cDNA of calpain from S. japonicum of two different isolates, Yamanashi strain (Sj-J) and Hunan strain (Sj-C), were 2, 468 bp and 2, 465 bp in length, including the same number (2, 274) of open reading frame. Nucleotide sequence and amino acid sequence between the two calpains are 99.1% and 98.8% identity, respectively. Sj-J and Sj-C calpains were considered to be translated as a preproenzyme, and a 746-amino acid mature enzyme contains eight motifs without a signal peptide at the N-terminal based on the deduced amino acid sequences. mRNA for calpain were detectable in different developmental stages, however, sera obtained from mice immunized with recombinant calpain showed enhanced binding to cercarial antigen. Human sera from S. japonicum-infected individuals recognized the large subunit of schistosomal calpain, and light-infected sera showed stronger reactivities to the recombinant calpain than moderate/high infection cases. When we tested synthetic peptides, there were four common human B cell epitopes in schistosomal calpain, all of which are shared with S. mansoni. Together with these results, calpain of S. japonicum seems to be not only a vaccine candidate, but also a target antigen for immunodiagnosis of human schistosomiasis.
Zhang
R
R
Department of Medical Zoology, Nagoya City University Medical School, 1 Azakawasumi, Mizuhocho, Mizuhoku, Nayoga 467-8601, Japan.
Suzuki
T
T
Takahashi
S
S
Yoshida
A
A
Kawaguchi
H
H
Maruyama
H
H
Yabu
Y
Y
Fu
J
J
Shirai
T
T
Ohta
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000032:analysis; D001483:Base Sequence; D015152:Blotting, Northern; D002154:Calpain / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D006801:Humans; D007114:Immunization; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D018510:RNA, Helminth / Q000737:chemistry / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012549:Schistosoma japonicum / Q000201:enzymology / Q000235:genetics*; D012552:Schistosomiasis / Q000517:prevention & control; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D000909:Antibodies, Helminth; D018509:DNA, Helminth; D000939:Epitopes; D018510:RNA, Helminth; D002154:Calpain