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One-Step Purification and Porin Transport Activity of the Major Outer Membrane Proteins P2 from Haemophilus influenzae, FomA from Fusobacterium nucleatum and PorB from Neisseria meningitidis

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Abstract

Bacterial porins are major outer membrane proteins that function as essential solute transporters between the bacteria and the extracellular environment. Structural features of porins are also recognized by eukaryotic cell receptors involved in innate and adaptive immunity. To better investigate the function of porins, proper refolding is necessary following purification from inclusion bodies [1, 2]. Using a single-step size exclusion chromatographic method, we have purified three major porins from pathogenic bacteria, the OmpP2 (P2) from Haemophilus influenzae, FomA from Fusobacterium nucleatum and PorB from Neisseria meningitidis, at high yield and report their unique solute transport activity with size exclusion limit. Furthermore, we have optimized their purification method and achieved improvement of their thermostability for facilitating functional and structural analyses.

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Acknowledgments

We thank Mika Jormakka (Centenary Institute, The University of Sydney) and Deana Toussi (Boston University) for critically reading the manuscript. We also thank Angelika Schierhorn (Martin Luther University) for mass spectrometry. This work was supported by the Bundesministerium für Bildung und Forschung (BMBF) ZIK program (FKZ 03Z2HN21) and ERDF (1241090001) (MT) by NIH grant R01 AI40944-01 (PM). The crystallographic data were tested at Swiss Light Source (SLS, Villingen) with support by the funding from the European Community's 7th Framework Programme (FP7/2007-2013) under BioStruct-X (grant agreement no. 283570, project ID BioStructx_5450). The authors declare no conflict of interest.

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Authors and Affiliations

  1. HALOmem, Membrane Protein Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Str.3, 06120, Halle (Saale), Germany

    Christof Kattner, Sabrina Pfennig & Mikio Tanabe

  2. Section of Infectious Diseases, Department of Medicine, Boston University School of Medicine, Boston, MA, USA

    Paola Massari

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  1. Christof Kattner
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Correspondence to Mikio Tanabe.

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Kattner, C., Pfennig, S., Massari, P. et al. One-Step Purification and Porin Transport Activity of the Major Outer Membrane Proteins P2 from Haemophilus influenzae, FomA from Fusobacterium nucleatum and PorB from Neisseria meningitidis . Appl Biochem Biotechnol 175, 2907–2915 (2015). https://doi.org/10.1007/s12010-014-1473-2

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