title: Direct Monitoring of β-Sheet Formation in the Outer Membrane Protein TtoA Assisted by TtOmp85.
abstract: Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was applied to investigate the folding of an outer membrane protein, TtoA, assisted by TtOmp85, both from the thermophilic eubacterium Thermus thermophilus. To directly monitor the formation of β-sheet structure in TtoA and to analyze the function of TtOmp85, we immobilized unfolded TtoA on an ATR crystal. Interaction with TtOmp85 initiated TtoA folding as shown by time-dependent spectra recorded during the folding process. Our ATR-FTIR experiments prove that TtOmp85 possesses specific functionality to assist β-sheet formation of TtoA. We demonstrate the potential of this spectroscopic approach to study the interaction of outer membrane proteins in vitro and in a time-resolved manner.
authors.affiliation: Department of Chemistry, ‡Department of Biology, and §Konstanz Research School Chemical Biology, University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Henke
authors.affiliation: Department of Chemistry, ‡Department of Biology, and §Konstanz Research School Chemical Biology, University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Wolfram
authors.initials: W
authors.lastname: Welte
authors.affiliation: Department of Chemistry, ‡Department of Biology, and §Konstanz Research School Chemical Biology, University of Konstanz , 78457 Konstanz, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Hauser
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry; D008958:Models, Molecular; D000072757:Protein Conformation, beta-Strand; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D016963:Thermus thermophilus / Q000737:chemistry*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D055767:Immobilized Proteins; D011994:Recombinant Proteins