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Expression, refolding, and initial structural characterization of the Y. pestis Ail outer membrane protein in lipids

Leigh A Plesniak1, Radhakrishnan Mahalakshmi, Candace Rypien

  • 1Sanford Burnham Medical Research Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA.

Biochimica Et Biophysica Acta
|October 2, 2010
PubMed

Insights

Ail, a key virulence factor of the plague bacterium Yersinia pestis, was structurally studied. Initial findings reveal Ail adopts a transmembrane beta-sheet structure in lipids, offering new therapeutic targets.

Area of Science:

  • Structural biology
  • Microbiology
  • Biochemistry

Background:

  • Ail is an outer membrane protein and virulence factor of Yersinia pestis, a Category A biothreat agent causing plague pandemics.
  • Ail facilitates bacterial adhesion and resistance to host defenses, making it a critical target for anti-plague therapies.
  • Limited knowledge exists regarding Ail's molecular functions, host interactions, and its three-dimensional structure.

Purpose of the Study:

  • To elucidate the molecular structure of Ail, an outer membrane protein of Yersinia pestis.
  • To prepare Ail for nuclear magnetic resonance (NMR) structural studies in lipid environments.

Main Methods:

  • Recombinant expression and purification of Ail.
  • Refolding and sample preparation for solution and solid-state NMR.
  • Structural studies using NMR and circular dichroism (CD) in lipid micelles and bilayers.

Main Results:

  • Successful recombinant expression, purification, and refolding of Ail were achieved.
  • Initial NMR and CD spectra indicate Ail adopts a defined transmembrane beta-sheet conformation within lipid environments.
  • The study provides the foundation for detailed structural and functional analysis of Ail.

Conclusions:

  • Ail forms a stable transmembrane beta-sheet structure in lipid bilayers, crucial for its function.
  • Understanding Ail's structure is vital for developing novel anti-plague therapeutics.
  • This work lays the groundwork for future investigations into Ail's role in Yersinia pestis pathogenesis.

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