Cloning and characterization of the SmIMP25 integral membrane protein of the parasitic helminth Schistosoma mansoni

Abstract

The cDNA and genomic clones encoding a 25 kDa integral membrane protein, termed SmIMP25, were isolated from Schistosoma mansoni. The 2.2 kb SmIMP25 mRNA was found in all developmental stages of the parasite tested: miracidium, sporocyst, cercaria and adult worm. The SmIMP25 gene is at least 16 kb long and it is split by four introns ranging in size from 36 bp to ≥ 9 kb. Excluding the introns, the gene and the cDNA show 100% sequence identity. The cDNA has an open reading frame encoding a protein 223 amino acids long. The predicted sequence reveals a distinct hydrophobic domain of 20 amino acids located 12 residues from the carboxyl-terminal end. The properties of this domain (marked hydrophobicity, size, flanking by charged residues and C-terminal location) are typical of the transmembrane segments of integral membrane proteins. The presence of three potential N-glycosylation sites is also consistent with membrane proteins that are often glycosylated at the extracellular domain. Accordingly we propose that SmIMP25 is an integral membrane protein in which residues 1–191 are extracellular, residues 192–211 comprise the hydrophobic domain that spans the membrane, and residues 212–223 are intracellular. The SmIMP25 was synthesized as a fusion protein in bacteria and antibodies were elicited in rabbits. Antibodies against SmIMP25 specifically precipitated a 25 kDa protein from cell-free products programmed by schistosome mRNA, in agreement with the size of the protein predicted from the cDNA sequence. Immunofluorescence studies showed SmIMP25 on the surface of the parasite. Surface molecules expressed at the host-parasite interface are likely to provide information on host parasite relationship and may serve as targets for protective immunity.