Protein-Lipid Interactions with Fusobacterium nucleatum Major Outer Membrane Protein FomA : Spin-Label EPR and Polarized Infrared Spectroscopy

Kleinschmidt, Jörg; Marsh, Derek; Vijay, N.; Anbazhagan, Veerappan

Protein-Lipid Interactions with Fusobacterium nucleatum Major Outer Membrane Protein FomA : Spin-Label EPR and Polarized Infrared Spectroscopy

Dateien

FomA_413_Biochem2008_Anbazhagan_etalFomA.pdfGröße: 1.63 MBDownloads: 385

Datum

2008

Autor:innen

Kleinschmidt, Jörg

Marsh, Derek

Vijay, N.

Anbazhagan, Veerappan

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Biologie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Biochemistry. 2008, 47(37), pp. 8414-8423. ISSN 0006-2960. eISSN 1520-4995. Available under: doi: 10.1021/bi800750s

Zusammenfassung

FomA, the major outer membrane protein of Fusobacterium nucleatum, was expressed and purified in Escherichia coli and reconstituted from detergent in bilayer membranes of phosphatidylcholines with chain lengths from C(12:0) to C(17:0). The conformation and orientation of membrane-incorporated FomA were determined from polarized, attenuated total reflection, infrared (IR) spectroscopy, and lipid-protein interactions with FomA were characterized by using electron paramagnetic resonance (EPR) spectroscopy of spin-labeled lipids. Approximately 190 residues of membranous FomA are estimated to be in a !-sheet configuration from IR band fitting, which is consistent with a 14-strand transmembrane !-barrel structure. IR dichroism of FomA indicates that the !-strands are tilted by !45° relative to the sheet/barrel axis and that the order parameter of the latter displays a discontinuity corresponding to hydrophobic matching with fluid C(13:0) lipid chains. The stoichiometry (Nb ) 23 lipids/monomer) of lipid-protein interaction from EPR demonstrates that FomA is not trimeric in membranes of diC(14:0) phosphatidylcholine and is consistent with a monomeric !-barrel of 14-16 strands. The pronounced selectivity of interaction found with anionic spin-labeled lipids places basic residues of the protein in the vicinity of the polar-apolar membrane interfaces, consistent with current topology models. Comparison with similar data from the 8- to 22-stranded E. coli outer membrane proteins, OmpA, OmpG, and FhuA, supports the above conclusions.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

KLEINSCHMIDT, Jörg, Derek MARSH, N. VIJAY, Veerappan ANBAZHAGAN, 2008. Protein-Lipid Interactions with Fusobacterium nucleatum Major Outer Membrane Protein FomA : Spin-Label EPR and Polarized Infrared Spectroscopy. In: Biochemistry. 2008, 47(37), pp. 8414-8423. ISSN 0006-2960. eISSN 1520-4995. Available under: doi: 10.1021/bi800750s

BibTex

@article{Kleinschmidt2008Prote-8118,
  year={2008},
  doi={10.1021/bi800750s},
  title={Protein-Lipid Interactions with Fusobacterium nucleatum Major Outer Membrane Protein FomA : Spin-Label EPR and Polarized Infrared Spectroscopy},
  number={37},
  volume={47},
  issn={0006-2960},
  journal={Biochemistry},
  pages={8414--8423},
  author={Kleinschmidt, Jörg and Marsh, Derek and Vijay, N. and Anbazhagan, Veerappan}
}

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