Molecular Cloning and Expression of a Gene EncodingCryptosporidium parvumGlycoproteins gp40 and gp15

Cryptosporidium parvum is a significant cause of diarrheal disease worldwide. The specific molecules that mediateC. parvum -host cell interactions and the molecular mechanisms involved in the pathogenesis of cryptosporidiosis are unknown. In this study we have shown that gp40, a mucin-like glycoprotein, is localized to the surface and apical region of invasive stages of the parasite and is shed from its surface. gp40-specific antibodies neutralize infection in vitro, and native gp40 binds specifically to host cells, implicating this glycoprotein inC. parvum attachment to and invasion of host cells. We have cloned and sequenced a gene designatedCpgp40/15 that encodes gp40 as well as gp15, an antigenically distinct, surface glycoprotein also implicated inC. parvum -host cell interactions. Analysis of the deduced amino acid sequence of the 981-bpCpgp40/15 revealed the presence of an N-terminal signal peptide, a polyserine domain, multiple predictedO -glycosylation sites, a single potentialN -glycosylation site, and a hydrophobic region at the C terminus, a finding consistent with what is required for the addition of a GPI anchor. There is a single copy ofCpgp40/15 in theC. parvum genome, and this gene does not contain introns. Our data indicate that the twoCpgp40/15 -encoded proteins, gp40 and gp15, are products of proteolytic cleavage of a 49-kDa precursor protein which is expressed in intracellular stages of the parasite. The surface localization of gp40 and gp15 and their involvement in the host-parasite interaction suggest that either or both of these glycoproteins may serve as effective targets for specific preventive or therapeutic measures for cryptosporidiosis.