Cryptosporidium p30, a galactose/N-acetylgalactosamine-specific lectin, mediates infection in vitro.
Cryptosporidium sp. cause human and animal diarrheal disease worldwide. The molecular mechanisms underlying Cryptosporidium attachment to, and invasion of, host cells are poorly understood. Previously, we described a surface-associated Gal/GalNAc-specific lectin activity in sporozoites of Cryptosporidium parvum. Here we describe p30, a 30-kDa Gal/GalNAc-specific lectin isolated from C. parvum and Cryptosporidium hominis sporozoites by Gal-affinity chromatography. p30 is encoded by a single copy gene containing a 906-bp open reading frame, the deduced amino acid sequence of which predicts a 302-amino acid, 31.8-kDa protein with a 22-amino acid N-terminal signal sequence. The p30 gene is expressed at 24-72 h after infection of intestinal epithelial cells. Antisera to recombinant p30 expressed in Escherichia coli react with an approximately 30-kDa protein in C. parvum and C. hominis. p30 is localized to the apical region of sporozoites and is predominantly intracellular in both sporozoites and intracellular stages of the parasite. p30 associates with gp900 and gp40, Gal/GalNAc-containing mucin-like glycoproteins that are also implicated in mediating infection. Native and recombinant p30 bind to Caco-2A cells in a dose-dependent, saturable, and Gal-inhibitable manner. Recombinant p30 inhibits C. parvum attachment to and infection of Caco-2A cells, whereas antisera to the recombinant protein also inhibit infection. Taken together, these findings suggest that p30 mediates C. parvum infection in vitro and raise the possibility that this protein may serve as a target for intervention.
Bhat
Najma
N
Division of Geographic Medicine and Infectious Diseases, Tufts-New England Medical Center, Boston, Massachusetts 02111, USA.
Joe
Angela
A
PereiraPerrin
Mercio
M
Ward
Honorine D
HD
D000116:Acetylgalactosamine / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D018938:Caco-2 Cells; D002846:Chromatography, Affinity; D003457:Cryptosporidiosis / Q000378:metabolism / Q000469:parasitology*; D003458:Cryptosporidium / Q000378:metabolism*; D016785:Cryptosporidium parvum / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D005690:Galactose / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry; D016366:Open Reading Frames; D034101:Sporozoites / Q000378:metabolism
D006023:Glycoproteins; D037102:Lectins; D009005:Monosaccharides; D000116:Acetylgalactosamine; D005690:Galactose