Identification of surface proteins on viable Plasmodium knowlesi merozoites
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Abstract:
Viable merozoites of Plasmodium knowlesi were isolated and the proteins that were labeled on intact merozoites by lactoperoxidase-catalyzed radioiodination were identified. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography of Triton soluble extracts of labeled merozoites demonstrated eight major bands ranging in apparent molecular weight from 150,000 D to 22,000 D. Exposure of intact merozoites to trypsin (10 microgram/ml) for 10 min resulted in the loss of the two highest molecular weight proteins (150,000 D and 105,000 D) and the appearance of two new bands at 70,000 D and 62,000 D. Trypsin treatment under these conditions also removed the receptor(s) for merozoite attachment to erythrocytes. Therefore, these high molecular weight proteins are candidates for the merozoite component that attaches to erythrocytes. There was no evidence that the labeled membrane components were serum or erythrocyte membrane components, two potential contaminants in the preparation. Anti-rhesus erythrocyte antibody did not precipitate labeled merozoite proteins. Furthermore, the immunoprecipitation of labeled merozoite proteins by rhesus anti-merozoite serum was not inhibited by erythrocyte ghosts.
Insights
High molecular weight proteins on Plasmodium knowlesi merozoites are key to erythrocyte attachment. These proteins are shed upon trypsin treatment, which also blocks merozoite invasion.
Area of Science:
- Malariology
- Parasitology
- Molecular Biology
Background:
- Plasmodium knowlesi merozoites infect erythrocytes, causing malaria.
- Identifying merozoite surface proteins involved in host cell invasion is crucial for understanding malaria pathogenesis.
Purpose of the Study:
- To identify Plasmodium knowlesi merozoite surface proteins responsible for erythrocyte attachment.
- To investigate the role of high molecular weight proteins in merozoite invasion.
Main Methods:
- Lactoperoxidase-catalyzed radioiodination of intact Plasmodium knowlesi merozoites.
- Sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography.
- Proteolytic treatment with trypsin and immunoprecipitation assays.
Main Results:
- Eight major merozoite proteins (150,000 D to 22,000 D) were identified.
- Trypsin treatment removed high molecular weight proteins (150,000 D and 105,000 D) and abolished erythrocyte attachment.
- No evidence of erythrocyte or serum protein contamination was found.
Conclusions:
- High molecular weight surface proteins of Plasmodium knowlesi merozoites are critical for erythrocyte binding.
- These proteins are potential targets for antimalarial drug development.