Crystal Structure of a Major Outer Membrane Protein from Thermus thermophilus HB27

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2009
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Journal of Molecular Biology. 2009, 385(5), pp. 1445-1455. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2008.12.003
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The thermophilic eubacterium Thermus thermophilus belongs to one of the oldest branches of evolution and has a multilayered cell envelope that differs from that of modern Gram-negative bacteria. Its outer membrane contains integral outer membrane proteins (OMPs), of which only a few are characterized. TtoA, a new p-barrel OMP, was identified by searching the genome sequence of strain HB27 for the presence of a C-terminal signature sequence. The structure of TtoA was determined to a resolution of 2.8 Å, representing the first crystal structure of an OMP from a thermophilic bacterium. TtoA consists of an eight-stranded p-barrel with a large extracellular part to which a divalent cation is bound. A five-stranded extracellular β-sheet protrudes out of the membrane-embedded transmembrane barrel and is stabilized by a disulfide bridge. The edge of this β-sheet forms crystal contacts that could mimic interactions with other proteins. In modem Gram-negative bacteria, the C-terminal signature sequence of OMPs is required for binding to an Omp85 family protein as a prerequisite for its assembly. We present hints that a similar assembly pathway exists in T. thermophilus by an in vitro binding assay, where unfolded TtoA binds to the Thermus Omp85 family protein TtOmp85, while a mutant without the signature sequence does not.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie
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outer membrane biogenesis, thermophile, T. thermophilus, TTC0834, TTC1475
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ISO 690BROSIG, Alexander, Jutta NESPER, Winfried BOOS, Wolfram WELTE, Kay DIEDERICHS, 2009. Crystal Structure of a Major Outer Membrane Protein from Thermus thermophilus HB27. In: Journal of Molecular Biology. 2009, 385(5), pp. 1445-1455. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2008.12.003
BibTex
@article{Brosig2009-02-06Cryst-1125,
  year={2009},
  doi={10.1016/j.jmb.2008.12.003},
  title={Crystal Structure of a Major Outer Membrane Protein from Thermus thermophilus HB27},
  number={5},
  volume={385},
  issn={0022-2836},
  journal={Journal of Molecular Biology},
  pages={1445--1455},
  author={Brosig, Alexander and Nesper, Jutta and Boos, Winfried and Welte, Wolfram and Diederichs, Kay}
}
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    <dcterms:abstract xml:lang="eng">The thermophilic eubacterium Thermus thermophilus belongs to one of the oldest branches of evolution and has a multilayered cell envelope that differs from that of modern Gram-negative bacteria. Its outer membrane contains integral outer membrane proteins (OMPs), of which only a few are characterized. TtoA, a new p-barrel OMP, was identified by searching the genome sequence of strain HB27 for the presence of a C-terminal signature sequence. The structure of TtoA was determined to a resolution of 2.8 Å, representing the first crystal structure of an OMP from a thermophilic bacterium. TtoA consists of an eight-stranded p-barrel with a large extracellular part to which a divalent cation is bound. A five-stranded extracellular β-sheet protrudes out of the membrane-embedded transmembrane barrel and is stabilized by a disulfide bridge. The edge of this β-sheet forms crystal contacts that could mimic interactions with other proteins. In modem Gram-negative bacteria, the C-terminal signature sequence of OMPs is required for binding to an Omp85 family protein as a prerequisite for its assembly. We present hints that a similar assembly pathway exists in T. thermophilus by an in vitro binding assay, where unfolded TtoA binds to the Thermus Omp85 family protein TtOmp85, while a mutant without the signature sequence does not.</dcterms:abstract>
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