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Molecular analysis of astacin-like metalloproteases of Ostertagia ostertagi.
- Source :
-
Parasitology [Parasitology] 2005 Jan; Vol. 130 (Pt 1), pp. 89-98. - Publication Year :
- 2005
-
Abstract
- In this study, we describe the molecular analysis of zinc-metalloproteases from the abomasal nematode Ostertagia ostertagi which were exclusively recognized by local antibodies of immune cattle. Full-length or partial coding sequences of 4 different zinc-metalloprotease cDNAs of Ostertagia (met-1, -2, -3 and -4) were amplified using gene-specific primers using the 3'- and 5'-Rapid Amplification of cDNA Ends (RACE) technique. Sequence analysis identified the cDNAs as encoding zinc-metalloproteases, which showed between 62% and 70% homology to a metalloprotease 1 precursor of Ancylostoma caninum. The full-length cDNA of met-1 consists of an open reading frame (ORF) of 586 amino acids which contains 5 potential N-glycosylation sites and a predicted zinc-binding domain (HEBXHXBGFXHEXXRXDRD). The complete coding sequence of met-3 contains an ORF of 508 aa and the same conserved zinc-binding domain. These domains are signature sequences of the astacin family of the superfamily of metzincin metalloproteases. The presence of a threonine amino acid after the third histidine in MET-1 and MET-3, however, may place them in a new family or subfamily. Real-time PCR analysis of L3, exsheathed L3, L4 and adult cDNA identified transcription of the 4 metalloproteases in different life-stages. The protein MET-1 was expressed in insect cells using the baculovirus expression system but the immunization of calves with this molecule did not lead to protection against challenge infection.
- Subjects :
- Amino Acid Sequence
Animals
Cattle
Cattle Diseases parasitology
Cattle Diseases prevention & control
Feces parasitology
Gene Expression Regulation, Developmental
Gene Expression Regulation, Enzymologic
Metalloendopeptidases
Metalloproteases immunology
Metalloproteases metabolism
Molecular Sequence Data
Ostertagia genetics
Ostertagiasis prevention & control
Ostertagiasis veterinary
Parasite Egg Count
Phylogeny
Sequence Alignment
Sequence Homology, Amino Acid
Vaccines
Metalloproteases chemistry
Ostertagia enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 0031-1820
- Volume :
- 130
- Issue :
- Pt 1
- Database :
- MEDLINE
- Journal :
- Parasitology
- Publication Type :
- Academic Journal
- Accession number :
- 15700760
- Full Text :
- https://doi.org/10.1017/s0031182004006274