title: 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.
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.
authors.affiliation: HALOmem, Membrane Protein Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Str.3, 06120, Halle (Saale), Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Kattner
authors.forename: Sabrina
authors.initials: S
authors.lastname: Pfennig
authors.forename: Paola
authors.initials: P
authors.lastname: Massari
authors.forename: Mikio
authors.initials: M
authors.lastname: Tanabe
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001692:Biological Transport; D002850:Chromatography, Gel / Q000379:methods*; D016967:Fusobacterium nucleatum / Q000737:chemistry / Q000378:metabolism; D006193:Haemophilus influenzae / Q000737:chemistry* / Q000378:metabolism; D009345:Neisseria meningitidis / Q000737:chemistry* / Q000378:metabolism; D018272:Porins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D055550:Protein Stability
chemical_list: D001425:Bacterial Outer Membrane Proteins; D018272:Porins