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Protein synthesis by a cell-free preparation from the bird malaria, Plasmodium lophurae
Abstract:
Cytoplasmic polyribosomes were isolated from the avian malaria parasite Plasmodium lophurae by lysis with 0.15% Triton X-100 followed by high speed centrifugation through a discontinuous sucrose gradient. Polyribosomes were protected from nuclease degradation using 100 mug/ml heparin or 50 mug/ml dextran sulfate. Cell-free incorporation of radioisotope-labeled amino acids required a pH 5 fraction (duck reticulocyte), Mg2+, and an energy-generating system. The protein synthesizing system was stimulated by the addition of polyuridylic acid. Optimum conditions for protein synthesis by the plasmodial system are described. The effects of drugs on the cell-free protein synthesizing system using duck reticulocyte and plasmodial ribosomes are reported.
Insights
Researchers isolated and characterized protein synthesis machinery from Plasmodium lophurae, an avian malaria parasite. This cell-free system, using duck reticulocyte components, enables study of parasite protein synthesis and drug effects.
Area of Science:
- Parasitology
- Molecular Biology
- Biochemistry
Background:
- Plasmodium lophurae is an avian malaria parasite.
- Understanding parasite protein synthesis is crucial for developing antimalarial strategies.
Purpose of the Study:
- To isolate and characterize cytoplasmic polyribosomes from Plasmodium lophurae.
- To establish a cell-free protein synthesizing system for studying parasite molecular mechanisms.
- To investigate the effects of drugs on this system.
Main Methods:
- Parasite lysis using Triton X-100.
- Isolation of polyribosomes via sucrose gradient centrifugation.
- Cell-free protein synthesis using radioisotope-labeled amino acids.
- Utilizing a pH 5 fraction from duck reticulocytes.
Main Results:
- Successfully isolated functional polyribosomes from Plasmodium lophurae.
- Identified requirements for cell-free protein synthesis: pH 5 fraction, Mg2+, and energy system.
- Demonstrated stimulation of protein synthesis by polyuridylic acid.
- Reported optimal conditions and drug effects on the system.
Conclusions:
- A functional cell-free protein synthesizing system was established for Plasmodium lophurae.
- This system provides a valuable tool for studying parasite-specific protein synthesis.
- The system can be used to screen for potential antimalarial drugs targeting protein synthesis.