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Food vacuole associated enolase in plasmodium undergoes multiple post-translational modifications: evidence for

Saudamini Shevade1, Nitin Jindal, Sneha Dutta

  • 1Department of Biological Sciences, Tata Institute of Fundamental Research, Colaba, Mumbai, India.

Plos One
|September 7, 2013
PubMed

Insights

Plasmodium enolase exhibits diverse sub-cellular localization and moonlighting functions, driven by post-translational modifications (PTMs). Ubiquitination and other PTMs on food vacuole-associated enolase explain its varied molecular weights and functions.

Area of Science:

  • Molecular Parasitology
  • Protein Biochemistry
  • Cell Biology

Background:

  • Plasmodium enolase localizes to multiple cellular compartments without specific targeting signals.
  • Post-translational modifications (PTMs) generate enolase variants with distinct organelle-specific localizations and potential moonlighting functions.

Purpose of the Study:

  • To investigate the nature and impact of PTMs on Plasmodium enolase, particularly variants associated with the food vacuole (FV).
  • To elucidate the specific PTMs, including ubiquitination, phosphorylation, and acetylation, responsible for the diverse forms and localization of FV-associated enolase.

Main Methods:

  • Immuno-precipitation and Western blot analysis to detect ubiquitinated forms of FV-associated enolase.
  • Mass spectrometry (MS) to identify and characterize PTMs on enolase variants.
  • Analysis of cytosolic and cytoskeleton-associated enolase PTMs in Plasmodium species.

Main Results:

  • FV-associated enolase exists in ~50, 65, and 75 kDa forms, with the latter two being ubiquitinated.
  • MS confirmed ubiquitination at K147, phosphorylation at Y148, and acetylation at K142/K384 on FV enolase.
  • Tri-ubiquitination via K147 was observed, with specific linear and atypical isopeptide linkages.
  • Phosphorylation sites were identified in cytosolic (S42) and cytoskeleton-associated (T337) enolase.

Conclusions:

  • PTMs, especially ubiquitination, are critical for the diverse localization and functions of Plasmodium enolase.
  • Specific ubiquitination patterns and other PTMs dictate the molecular weight and cellular association of enolase variants.
  • The study provides detailed insights into the PTM landscape of Plasmodium enolase and its FV proteome interactions.

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