Identification of a coronin-like protein in Babesia species.
The present study was designed to immunochemically identify a coronin-like protein in Babesia bovis, B. bigemina, B. divergens, and B. canis. A 2-kbp cDNA insert of B. bovis carried by plasmid BvN9 was sequenced by the dideoxichain-termination method on both strands. The cDNA insert contained a 1719-bp long open reading frame coding for a deduced protein sequence of 61.7 kDa. Sequence analysis using the PSI-BLAST program revealed about 30% protein sequence identity with a coronin-like protein of Plasmodium falciparum. The encoding sequence of the cDNA insert lacking 70 amino acids at the N-terminal was subcloned in frame into pGEX 4T-3 to produce a recombinant glutathione S-transferase (GST)-pBv fusion protein. Polyclonal antibodies prepared in rabbits immunized with the purified GST-fusion protein recognized a Babesia-specific component of approximately 60 kDa by immunoprecipitation with [35S]methionine-labeled parasites. However, two molecules with relative sizes of 60 and 70 kDa were recognized in Babesia-infected erythrocyte extracts by immunobloting analysis. The 70-kDa component was apparently of host erythrocyte origin. In an indirect fluorescent antibody test, the rabbit serum strongly reacted with the merozoite stage of the four Babesia species, but also, although weakly, with the host erythrocyte. A cosedimentation assay performed with GST-pBv fusion protein and exogenous actin from rabbit liver showed that the GST-pBv fusion protein, but not the GST protein, was associated to actin. From these results, we conclude that the protein present in the four Babesia species analyzed here may be considered as a novel coronin-like, actin-binding protein.
Figueroa
Julio V
JV
CENID-PAVET, INIFAP, Juitepec, Morelos, Mexico. figueroa.julio@inifap.gob.mx
Precigout
Eric
E
Carcy
Bernard
B
Gorenflot
André
A
D000595:Amino Acid Sequence; D000818:Animals; D016793:Babesia bovis / Q000737:chemistry*; D001404:Babesiosis; D002417:Cattle; D002418:Cattle Diseases; D018076:DNA, Complementary / Q000032:analysis; D004912:Erythrocytes; D015151:Immunoblotting; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D008840:Microfilament Proteins