Protein analysis of Babesia caballi merozoites by two-dimensional polyacrylamide gel electrophoresis and western blotting.
Abstract: Babesia caballi merozoites were prepared by combining two improved methods of cultivation and purification of merozoites using Percoll-gradiation, and the protein compositions of merozoites were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. The relative molecular masses of the major proteins and protein masses separated by electrophoresis were >94, 80-70, 50-45, 34-30, 30-28 and 18 kDa. By Western blotting, twelve proteins or protein groups were recognized by pooled sera from two horses experimentally infected with B. caballi. Among twelve proteins, five new proteins (54, 30-26, 24, and two 18 kDa) were identified, and the 48 kDa protein was revealed to consist of 2 components in the B. caballi merozoite. One protein (54 kDa) of B. caballi was also recognized by the pooled sera from two horses experimentally infected with B. equi.
Publication Date: 2000-04-19 PubMed ID: 10770608DOI: 10.1292/jvms.62.323Google Scholar: Lookup
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- Journal Article
Summary
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The research article focuses on a comprehensive analysis carried out on the protein composition of Babesia caballi merozoites, a type of parasite. The resilient protein compositions were identified using advanced techniques like two-dimensional polyacrylamide gel electrophoresis and Western blotting.
Methods Used in Research
The study employs advanced methods of cultivation and purification of merozoites. These include:
- Percoll-gradiation: This is a method used to purify cells, viruses, and subcellular organelles at low temperatures.
- Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE): This is used to separate proteins by mass and charge. It involves two steps. First is based on the protein charge and second is based on protein mass.
- Western blotting: This technique is used to detect specific proteins in a sample of tissue homogenate or extract.
Findings from the Study
The research identifies several proteins within the Babesia caballi merozoites:
- The major proteins and protein masses separated by electrophoresis had relative molecular masses of >94, 80-70, 50-45, 34-30, 30-28 and 18 kDa.
- Upon executing Western blotting, twelve different proteins or protein groups were recognized by pooled sera (blood serum) from two horses that were experimentally infected with the Babesia caballi.
- A notable finding is the identification of five new proteins, including two 18 kDa proteins. One particularly remarkable protein (54 kDa) was recognized both in B. caballi and by sera from horses experimentally infected with B. equi, another parasite species.
- Furthermore, it was revealed that the 48 kDa protein actually consists of two components in the B. caballi merozoite.
Significance of the Research
This research is significant as it:
- Offers deeper insight into the protein composition of B. caballi merozoites which may be crucial for developing interventions against it.
- Discovery of the common 54 kDa protein in B. caballi and B. equi suggests potential common targets for vaccine or drug development against these parasites.
- Identification of these proteins could also provide valuable information for future research into the biology and pathogenesis of these parasites.
Cite This Article
APA
Ikadai H, Kabamoto S, Xuan X, Igarashi I, Nagasawa H, Fujisaki K, Suzuki N, Mikami T.
(2000).
Protein analysis of Babesia caballi merozoites by two-dimensional polyacrylamide gel electrophoresis and western blotting.
J Vet Med Sci, 62(3), 323-327.
https://doi.org/10.1292/jvms.62.323 Publication
Researcher Affiliations
- The Research Center for Protozoan Molecular Immunology, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
MeSH Terms
- Animals
- Babesia / chemistry
- Babesiosis / parasitology
- Blotting, Western / veterinary
- Electrophoresis, Gel, Two-Dimensional / veterinary
- Horse Diseases / parasitology
- Horses
- Molecular Weight
- Protozoan Proteins / chemistry
Citations
This article has been cited 2 times.- Hirata H, Ikadai H, Yokoyama N, Xuan X, Fujisaki K, Suzuki N, Mikami T, Igarashi I. Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay. J Clin Microbiol 2002 Apr;40(4):1470-4.
- Ikadai H, Xuan X, Igarashi I, Tanaka S, Kanemaru T, Nagasawa H, Fujisaki K, Suzuki N, Mikami T. Cloning and expression of a 48-kilodalton Babesia caballi merozoite rhoptry protein and potential use of the recombinant antigen in an enzyme-linked immunosorbent assay. J Clin Microbiol 1999 Nov;37(11):3475-80.
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