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Expression of Pneumocystis jirovecii major surface glycoprotein in Saccharomyces cerevisiae

Geetha Kutty1, Katherine J England, Joseph A Kovacs

  • 1Critical Care Medicine Department, National Institutes of Health (NIH) Clinical Center, Bethesda, MD 20892, USA.

The Journal of Infectious Diseases
|March 28, 2013
PubMed

Insights

Researchers expressed the major surface glycoprotein (Msg) from Pneumocystis jirovecii in yeast. This engineered yeast exhibited enhanced adherence to lung cells, aiding Pneumocystis research.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • The major surface glycoprotein (Msg) is crucial for Pneumocystis organism attachment to host cells.
  • Understanding Msg's function requires effective expression and localization studies.

Purpose of the Study:

  • To express Pneumocystis jirovecii Msg in Saccharomyces cerevisiae.
  • To investigate methods for targeting recombinant Msg to the yeast cell wall for functional studies.

Main Methods:

  • Codon-optimized DNA construct for P. jirovecii Msg expression in yeast.
  • Engineering Msg fusion protein with yeast and fungal cell wall components for targeted localization.
  • Immunofluorescence and enzymatic assays to confirm cell wall localization.
  • Assessing yeast adherence to A549 alveolar epithelial cells.

Main Results:

  • Recombinant Msg initially localized to the yeast plasma membrane.
  • Engineered Msg fusion protein successfully localized to the yeast cell wall.
  • Yeast expressing Msg on its surface showed increased adherence to A549 cells.

Conclusions:

  • Heterologous expression of P. jirovecii Msg in yeast is feasible.
  • Targeting Msg to the yeast cell wall facilitates studies on its adhesive properties.
  • This system provides a valuable tool for investigating Pneumocystis Msg biology.
Keywords:
GPI anchored proteinPneumocystis jiroveciiantigenic variationmajor surface glycoproteinupstream conserved sequence

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