Journal of Protein Chemistry

, Volume 17, Issue 7, pp 719–728 | Cite as

Characterization of a subunit structure and stability of the recombinant porin fromNeisseria gonorrhoeae

  • Yury V. Matsuka
  • Deborah A. Dilts
  • Susan Hoiseth
  • Rasappa Arumugham


An outer membrane PIA protein fromNeisseria gonorrhoeae strain FA19 was expressed inEscherichia coli and refoldedin vitro in the presence of zwitterionic detergent. Its proper folding and subunit organization was confirmed by comparison with the native counterpart. The unfolding of PIA has been investigated using fluorescence spectroscopy and analytical size-exclusion chromatography methods. Analysis of the denaturation pathway of the PIA revealed that it forms an unusually labile quaternary structure. In the presence of 1 M guanidinium chloride (GdmCl) or upon heating up to 50°C, dissociation of the PIA oligomer was observed resulting in the formation of folded monomeric intermediates. Unfolding of monomers occurs at 80°C or in the presence of 4.3 M GdmCl, indicating high intrinsic stability toward both GdmCl and elevated temperatures. Both oligomeric and monomeric forms of PIA exhibited affinity to the hydrophobic probe 1-anilinonaphthalene-8-sulfonic acid (ANS) and bind withK d=80 and 130 µM, respectively. Denaturation of the PIA completely abolished affinity to ANS, suggesting that hydrophobicity is a property of the folded state of the porin.

Key words

N. gonorrhoeae porin expression unfolding ANS intermediate state 


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Copyright information

© Plenum Publishing Corporation 1998

Authors and Affiliations

  • Yury V. Matsuka
    • 1
  • Deborah A. Dilts
    • 2
  • Susan Hoiseth
    • 2
  • Rasappa Arumugham
    • 1
  1. 1.Department of Protein and Analytical ChemistryWyeth-Lederle Vaccines and PediatricsWest Henrietta
  2. 2.Department of Molecular BiologyWyeth-Lederle Vaccines and PediatricsWest Henrietta

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