Identification and characterization of myophilin, a muscle-specific antigen of Echinococcus granulosus.
A muscle-specific gene of Echinococcus granulosus has been identified and characterized. A lambda gt11 clone (10P1), containing an incomplete copy of the gene, was originally isolated from a larval E. granulosus cDNA library by serum antibodies from dogs infected with the parasite. The full-length cDNA sequence was obtained by PCR amplification of cDNA from an adult E. granulosus lambda gt22A library. Southern blot analysis indicated the presence of the gene as a single copy in the genome of E. granulosus and also detected homologous genes in genomic DNA of E. multilocularis and Taenia saginata. The 21.2-kDa protein deduced from the complete cDNA sequence contains two regions of 12 amino acids with similarity to the EF-hand motif of calcium binding proteins. Antibodies raised against the purified 10P1-GST fusion protein detected a 22-kDa antigen in the E. granulosus developmental stages examined. Immunoelectron microscopy localized the native protein in the muscle of the parasite. The amino-acid sequence of the E. granulosus protein shows significant homology to the muscle proteins mp20 of Drosophila melanogaster, chicken SM22 alpha and mammalian calponin, and also to the neuronal protein NP25 of rats. A conserved carboxy-terminal motif of 17 amino acids is present in all the homologous proteins and is proposed to be the characteristic feature of a novel protein family. The term myophilin is proposed for the E. granulosus protein due to its localization and homology to other muscle proteins.
Martin
R M
RM
University of Melbourne, Department of Veterinary Science, Werribee, Victoria, Australia.
Gasser
R B
RB
Jones
M K
MK
Lightowlers
M W
MW
D000595:Amino Acid Sequence; D000818:Animals; D000835:Animals, Wild / Q000469:parasitology; D000947:Antigens, Helminth; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000737:chemistry; D003001:Cloning, Molecular; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D004443:Echinococcosis / Q000469:parasitology / Q000662:veterinary; D004446:Echinococcus / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D007814:Larva; D008840:Microfilament Proteins; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D000096985:Calponins
D000947:Antigens, Helminth; D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D015801:Helminth Proteins; D008840:Microfilament Proteins; D009124:Muscle Proteins; D011993:Recombinant Fusion Proteins; D000096985:Calponins; C095050:myophilin protein, Echinococcus granulosus