Stability of the Thermus thermophilus outer membrane protein TtoA against heat and denaturants

Article type: Research Article

Authors: Henke, Katharina | Odermatt, Meike | Welte, Wolfram | Hauser, Karin^;

Affiliations: Department of Chemistry, University of Konstanz, Konstanz, Germany | Department of Biology, University of Konstanz, Konstanz, Germany

Note: [] Corresponding author: Karin Hauser, Department of Chemistry, University of Konstanz, 78457 Konstanz, Germany. Tel.: +49 7531 88 5356; Fax: +49 7531 88 3139; E-mail: [email protected]

Abstract: TtoA is a major outer membrane protein (Omp) with β-barrel structure from the thermophilic eubacterium T. thermophilus. FT-IR spectroscopy and SDS PAGE analysis were used to monitor the stability of detergent-solubilised TtoA under denaturing conditions. Heat as well as the common denaturants urea and SDS were applied to affect the TtoA structure. The protein has proven to be extremely thermostable in its native form. Denaturants likewise only have little effects on the protein's structure. In detail, a SDS concentration of 1% as is typical for Laemmli buffer does not have an impact on the structure of TtoA, even in combination with a temperature of 99°C. The same holds true for urea concentrations below 8 M. Only the combination of highly concentrated urea or SDS in combination with incubating the protein at 99°C for 10 minutes leads to a change of the secondary structure in TtoA.

Keywords: β-barrel, infrared spectroscopy, stability, outer membrane protein

DOI: 10.3233/BSI-140063

Journal: Biomedical Spectroscopy and Imaging, vol. 3, no. 1, pp. 51-56, 2014