The N-terminal segment of glyceraldehyde-3-phosphate dehydrogenase of Haemonchus contortus interacts with complements C1q and C3.
Haemonchus contortus, an economically important blood-sucking parasite of sheep and goats, survives the harsh host gut environment by secreting a number of proteins referred as excretory/secretory (ES) products. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme, is one of the components of H. contortus ES products. The parasite enzyme binds to complement C3 and inhibits its activity. In this study, the C3-binding activity of the parasite GAPDH was mapped to the N-terminal part of the enzyme by generating defined recombinant fragments of the protein. The N-terminal fragment also trapped complement C1q but not C5 and inhibited complement-mediated lysis of sensitized sheep erythrocytes. Competitive binding assay indicates different binding regions for C1q and C3 proteins. GAPDH stimulated proliferation of goat peripheral blood mononuclear cells in vitro and reacted with the sera from H. contortus-infected animals. However, the fragments of GAPDH did not stimulate cell proliferation nor reacted with the infected animal sera. Furthermore, denatured GAPDH failed to react with the infected animal sera in dot blot suggesting conformation-dependent epitope. These results demonstrate an elegant strategy of the parasite to completely shut down complement activation and identify GAPDH as a promising target for future therapeutic intervention.
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, India.
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Vedamurthy
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, India.
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Sahoo
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, India.
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Devi
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, India.
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Murugavel
Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar, India.
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Joshi
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049109:Cell Proliferation; D003167:Complement Activation; D015922:Complement C1q / Q000378:metabolism*; D003176:Complement C3 / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006041:Goats; D006188:Haemonchiasis / Q000469:parasitology / Q000662:veterinary; D006189:Haemonchus / Q000201:enzymology* / Q000276:immunology*; D007963:Leukocytes, Mononuclear; D008969:Molecular Sequence Data; D011817:Rabbits; D012756:Sheep
D003176:Complement C3; D015922:Complement C1q; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
conformation dependent epitope; excretory/secretory products; glyceraldehyde-3-phosphate dehydrogenase; recombinant protein fragments