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Euglena gracilis chloroplast elongation factor Tu. Purification and initial characterization.

S P Sreedharan, C M Beck, L L Spremulli

    The Journal of Biological Chemistry
    |March 10, 1985
    PubMed
    Summary

    The chloroplast protein synthesis elongation factor Tu (EF-Tuchl) from Euglena gracilis was purified and characterized. This factor functions similarly to prokaryotic elongation factors, highlighting the link between chloroplast and bacterial translational systems.

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    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • Chloroplasts possess their own protein synthesis machinery, distinct from cytoplasmic systems.
    • Elongation factor Tu (EF-Tu) is crucial for protein synthesis in prokaryotes and organelles.
    • Understanding organellar EF-Tu provides insights into endosymbiotic relationships.

    Purpose of the Study:

    • To purify and characterize the chloroplast elongation factor Tu (EF-Tuchl) from Euglena gracilis.
    • To investigate the functional properties and interactions of EF-Tuchl.
    • To explore the evolutionary relationship between chloroplast and prokaryotic translational systems.

    Main Methods:

    • DEAE-Sephadex chromatography of Euglena postribosomal supernatant.
    • Gel filtration chromatography for complex isolation and molecular weight determination.

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  • In vitro activity assays on homologous, E. coli, and eukaryotic cytoplasmic ribosomes.

    • Main Results:

    • Two forms of EF-Tuchl were identified: a complex with a stimulatory factor and free EF-Tuchl.
    • Purified EF-Tuchl is a 50,000 MW polypeptide.
    • EF-Tuchl showed activity on E. coli ribosomes but not eukaryotic cytoplasmic ribosomes.
    • EF-Tuchl interacts with E. coli EF-Ts and is sensitive to N-ethylmaleimide and kirromycin.

    Conclusions:

    • Euglena chloroplast EF-Tuchl shares significant functional and structural similarities with prokaryotic EF-Tu.
    • These findings support the endosymbiotic origin of chloroplasts.
    • Chloroplast protein synthesis machinery retains prokaryotic characteristics.