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Cellular and Molecular Life Sciences

, Volume 74, Issue 12, pp 2319–2332 | Cite as

Crystal structure and biochemical characterization of the transmembrane PAP2 type phosphatidylglycerol phosphate phosphatase from Bacillus subtilis

  • Meriem El Ghachi
  • Nicole Howe
  • Rodolphe Auger
  • Alexandre Lambion
  • Annick Guiseppi
  • François Delbrassine
  • Guillaume Manat
  • Sophie Roure
  • Sabine Peslier
  • Eric Sauvage
  • Lutz Vogeley
  • Juan-Carlos Rengifo-Gonzalez
  • Paulette Charlier
  • Dominique Mengin-Lecreulx
  • Maryline Foglino
  • Thierry Touzé
  • Martin Caffrey
  • Frédéric Kerff
Original Article

Abstract

Type 2 phosphatidic acid phosphatases (PAP2s) can be either soluble or integral membrane enzymes. In bacteria, integral membrane PAP2s play major roles in the metabolisms of glycerophospholipids, undecaprenyl-phosphate (C55-P) lipid carrier and lipopolysaccharides. By in vivo functional experiments and biochemical characterization we show that the membrane PAP2 coded by the Bacillus subtilis yodM gene is the principal phosphatidylglycerol phosphate (PGP) phosphatase of B. subtilis. We also confirm that this enzyme, renamed bsPgpB, has a weaker activity on C55-PP. Moreover, we solved the crystal structure of bsPgpB at 2.25 Å resolution, with tungstate (a phosphate analog) in the active site. The structure reveals two lipid chains in the active site vicinity, allowing for PGP substrate modeling and molecular dynamic simulation. Site-directed mutagenesis confirmed the residues important for substrate specificity, providing a basis for predicting the lipids preferentially dephosphorylated by membrane PAP2s.

Keywords

Membrane protein structure Bacterial lipids metabolism Undecaprenyl phosphate Peptidoglycan-related lipid 

Notes

Acknowledgements

This work was supported by the Belgian program of Interuniversity Attraction Poles initiated by the Federal Office for Scientific Technical and Cultural Affairs (IAP no. P7/44), the FRS-FNRS (MIS F.4518.12, IISN 4.4503.11), the Tournesol/Hubert Curien partnership between Belgium and France (R.CFRA.1567), the Science Foundation Ireland (Grant Number 12/IA/1255), the Agence Nationale de la Recherche (Bactoprenyl project, ANR-11-BSV3-002), the Centre National de la Recherche Scientifique and the University of Paris-Sud (UMR 9198) and the Aix-Marseille University. The assistance and support of beamline scientists at the Advanced Photon Source (23-ID) are acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare no competing financial interests.

Supplementary material

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Supplementary material 1 (PDF 102 KB)
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Copyright information

© Springer International Publishing 2017

Authors and Affiliations

  • Meriem El Ghachi
    • 1
  • Nicole Howe
    • 2
  • Rodolphe Auger
    • 3
  • Alexandre Lambion
    • 1
  • Annick Guiseppi
    • 4
  • François Delbrassine
    • 1
  • Guillaume Manat
    • 3
  • Sophie Roure
    • 3
  • Sabine Peslier
    • 4
  • Eric Sauvage
    • 1
  • Lutz Vogeley
    • 2
  • Juan-Carlos Rengifo-Gonzalez
    • 1
  • Paulette Charlier
    • 1
  • Dominique Mengin-Lecreulx
    • 3
  • Maryline Foglino
    • 4
  • Thierry Touzé
    • 3
  • Martin Caffrey
    • 2
  • Frédéric Kerff
    • 1
  1. 1.Centre d’Ingénierie des Protéines, InBioSUniversité de LiègeLiègeBelgium
  2. 2.Membrane Structural and Functional Biology GroupSchools of Medicine and Biochemistry and Immunology, Trinity College DublinDublin 2Ireland
  3. 3.Institute for Integrative Biology of the Cell (I2BC), CEA, CNRSUniv Paris-Sud, Université Paris-SaclayGif-sur-Yvette cedexFrance
  4. 4.Laboratoire de Chimie Bactérienne UMR 7283Aix-Marseille UniversitéMarseilleFrance

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