The microneme protein MIC3 of Toxoplasma gondii is a secretory adhesin that binds to both the surface of the host cells and the surface of the parasite.
Assay of the adhesion of cultured cells on Toxoplasma gondii tachyzoite protein Western blots identified a major adhesive protein, that migrated at 90 kDa in non-reducing gels. This band comigrated with the previously described microneme protein MIC3. Cellular binding on Western blots was abolished by MIC3-specific monoclonal and polyclonal antibodies. The MIC3 protein affinity purified from tachyzoite lysates bound to the surface of putative host cells. In addition, T. gondii tachyzoites also bound to immobilized MIC3. Immunofluorescence analysis of T. gondii tachyzoite invasion showed that MIC3 was exocytosed and relocalized to the surface of the parasite during invasion. The cDNA encoding MIC3 and the corresponding gene have been cloned, allowing the determination of the complete coding sequence. The MIC3 sequence has been confirmed by affinity purification of the native protein and N-terminal sequencing. The deduced protein sequence contains five partially overlapping EGF-like domains and a chitin binding-like domain, which can be involved in protein-protein or protein-carbohydrate interactions. Taken together, these results suggest that MIC3 is a new microneme adhesin of T. gondii.
Garcia-Réguet
N
N
Biologie moléculaire et Pathogénèse des Sporozoaires, Institut de Biologie de Lille, France.
Lebrun
M
M
Fourmaux
M N
MN
Mercereau-Puijalon
O
O
Mann
T
T
Beckers
C J
CJ
Samyn
B
B
Van Beeumen
J
J
Bout
D
D
Dubremetz
J F
JF
D018829:Adhesins, Bacterial; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D002460:Cell Line / Q000378:metabolism / Q000469:parasitology; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D018503:Genome, Protozoan; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014122:Toxoplasma / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D018829:Adhesins, Bacterial; D002352:Carrier Proteins; C421382:MIC3 protein, Toxoplasma gondii; D015800:Protozoan Proteins