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. 2015 Mar;175(6):2907-15. doi: 10.1007/s12010-014-1473-2. Epub 2015 Jan 10.

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.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Outer Membrane Proteins / chemistry
  • Bacterial Outer Membrane Proteins / isolation & purification*
  • Bacterial Outer Membrane Proteins / metabolism
  • Biological Transport
  • Chromatography, Gel / methods*
  • Fusobacterium nucleatum / chemistry
  • Fusobacterium nucleatum / metabolism
  • Haemophilus influenzae / chemistry*
  • Haemophilus influenzae / metabolism
  • Neisseria meningitidis / chemistry*
  • Neisseria meningitidis / metabolism
  • Porins / chemistry
  • Porins / isolation & purification*
  • Porins / metabolism
  • Protein Stability

Substances

  • Bacterial Outer Membrane Proteins
  • Porins