Biochemical and Biophysical Research Communications
An extensively associated dimer in the structure of the C713S mutant of the TIR domain of human TLR2
Section snippets
Materials and methods
Cloning, protein expression, and purification. The C713S mutant of the TIR domain was prepared with the QuikChange mutagenesis kit (Stratagene) and sequenced to confirm the incorporation of the mutation. The mutant protein was expressed and purified using the same protocol as that for the wild-type protein [13].
Crystallization and data collection. Crystals of the C713S mutant were obtained at 21 °C by the hanging drop vapor diffusion method. The reservoir solution contained 100 mM MES (pH 6.5) or
Overall structure and conformational variability of the BB loop
The crystal structure of the C713S mutant of the TIR domain of human TLR2 has been determined at 3.2 Å resolution. The statistics for the structure refinement are summarized in Table 1. The atomic structure has been deposited at the Protein Data Bank (Accession No. 1077). There are five copies of the TIR domain molecule in the crystallographic asymmetric unit and they will be called molecules A through E here. In all these molecules, the first 13 residues of the protein (626–638) and a portion
Acknowledgements
We thank Kevin D’Amico and Steve Wassermann for access to the 32-ID beamline, Craig Ogata for access to the X4A beamline, Ming Li for helping to set up the TLR2 receptor assays, and John Sims for helpful discussions. This research was supported in part by a grant from the National Institutes of Health (AI49475 to L.T.).
References (22)
- et al.
Phylogenetic perspectives in innate immunity
Science
(1999) - et al.
Innate immunity: the virtues of a nonclonal system of recognition
Cell
(1997) - et al.
Toll-like receptors in the induction of the innate immune response
Nature
(2000) - et al.
Toll-like receptors: critical proteins linking innate and acquired immunity
Nat. Immunol.
(2001) - et al.
Toll-like receptors: key mediators of microbe detection
Curr. Opin. Immunol.
(2002) - et al.
Drosophila Toll and IL-1 receptor
Nature
(1991) - et al.
Dominant and recessive mutations define functional domains of Toll, a transmembrane protein required for dorsal–ventral polarity in the Drosophila embryo
Genes Dev.
(1991) - et al.
Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene
Science
(1998) - et al.
Cell activation and apoptosis by bacterial lipoproteins through Toll-like receptor-2
Science
(1999) - et al.
Toll-like receptor 2 functions as a pattern recognition receptor for diverse bacterial products
J. Biol. Chem.
(1999)
Differential roles of TLR2 and TLR4 in recognition of gram-negative and gram-positive bacterial cell wall components
Immunity
Cited by (99)
Characterization, expression and function analysis of the TLR3 gene in golden pompano (Trachinotus ovatus)
2021, Developmental and Comparative ImmunologyRegulation of signaling by cooperative assembly formation in mammalian innate immunity signalosomes by molecular mimics
2020, Seminars in Cell and Developmental BiologyStructure of a prokaryotic SEFIR domain reveals two novel SEFIR-SEFIR interaction modes
2018, Journal of Structural BiologyCitation Excerpt :Both SEFIR family and TIR family belong to the STIR superfamily, of which the members share similar α/β fold and are known to mediate physical protein-protein interactions in cytosolic signal transductions. Several TIR-TIR complex structures have been reported, including human, plant and microbial TIR domains (e.g. TLR2, TLR6, TLR10, MAL/TIRAP, IL-1RAPL, L6, RPS4, RRS1TcpB, PdTIR etc.) (Tao et al., 2002; Valkov et al., 2011; Lin et al., 2012; Nyman et al., 2008; Alaidarous et al., 2014; Jang and Park, 2014; Chan et al., 2010; Bernoux et al., 2011; Snyder et al., 2014; Khan et al., 2004). Structural analysis combined with biochemical data revealed that TIR domain forms various different kinds of dimers.
A polar SxxS motif drives assembly of the transmembrane domains of Toll-like receptor 4
2017, Biochimica et Biophysica Acta - BiomembranesThe molecular mechanisms of signaling by cooperative assembly formation in innate immunity pathways
2017, Molecular Immunology
- 1
Present address: Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA.