Characterization of Sm13, a tegumental antigen of Schistosoma mansoni.
Sm13, a 13-kDa Schistosoma mansoni tegumental antigen, is one of the principal polypeptides recognized by antibodies from mice protectively vaccinated with adult-worm tegumental membranes. To obtain the complete gene encoding Sm13 we subcloned and sequenced a cDNA and a fragment of a genomic clone. The collated sequence contains 1,088 nucleotides and represents the full-length open reading frame of the gene, encoding a protein of 104 amino acids with a calculated molecular mass of 11,923 Da, compatible with the native protein identified in the tegumental membranes. The sequence derived from genomic DNA contains a 45-nucleotide intron. The analysis of the predicted protein suggests the presence of both N- and C-terminal hydrophobic membrane-spanning segments, and the coding region contains no homology in the currently available data bases. Additionally, the coding region is preceded by putative CCAAT and TATA boxes that may be involved in the control of expression. Western-blot analysis and indirect immunofluorescence resulted in the identification of a 13-kDa protein (Sm13) in the tegument of adult worms. The present study reveals that Sm13 behaves as an integral membrane protein upon partitioning in Triton X-1 14 and that it is present in worms of 3 weeks or older but not in schistosomula or miracidia. Moreover, it is also specifically recognized by sera from some schistosomiasis patients in enzyme-linked immunosorbent assay and Western-blot analysis, suggesting that it is immunogenic in human schistosomiasis.
Abath
F G
FG
Centro de Pesquisas Aggeu Magalhães, Fundação, Cidade Universitária, Recife, Brazil. fabath@gene.dbbm.fiocruz.br
Xavier
E M
EM
Allen
R
R
Gomes
Y M
YM
Lucena-Silva
N
N
Baliza
M
M
Simpson
A J
AJ
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis / Q000235:genetics*; D000954:Antigens, Surface / Q000032:analysis / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D015801:Helminth Proteins; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018510:RNA, Helminth / Q000032:analysis; D012550:Schistosoma mansoni / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000947:Antigens, Helminth; D000954:Antigens, Surface; D018509:DNA, Helminth; D015801:Helminth Proteins; D008565:Membrane Proteins; D018510:RNA, Helminth; C419305:Sm13 antigen protein, Schistosoma mansoni