Projects per year
Abstract
As part of the mammalian innate immune response, Toll-like receptors 3 and 4 can signal via the adaptor protein TRIF/TICAM-1 to elicit the production of type-I interferons and cytokines. Recent studies have suggested an auto-inhibitory role for the N-terminal domain (NTD) of TRIF. This domain has no significant sequence similarity to proteins of known structure. In this paper, the crystallization and X-ray diffraction analysis of TRIF-NTD and its selenomethionine-labelled mutant TRIF-NTD(A66M/L113M) are reported. Thin plate-like crystals of native TRIF-NTD obtained using polyethylene glycol 3350 as precipitant diffracted X-rays to 1.9 A resolution. To facilitate phase determination, two additional methionines were incorporated into the protein at positions chosen based on the occurrence of methionines in TRIF homologues in different species. Crystals of the selenomethionine-labelled protein were obtained under conditions similar to the wild-type protein; these crystals diffracted X-rays to 2.5 A resolution. The TRIF-NTD and TRIF-NTD(A66M/L113M) crystals have the symmetry of space groups P212121 and P1, and most likely contain two and four molecules in the asymmetric unit, respectively. These results provide a sound foundation for the future structure determination of this novel domain.
Original language | English |
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Pages (from-to) | 766 - 770 |
Number of pages | 5 |
Journal | Acta Crystallographica Section F: Structural Biology Communications |
Volume | 69 |
Issue number | 7 |
DOIs | |
Publication status | Published - 2013 |
Projects
- 1 Finished
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Molecular and structural basis of signalling by TIR domain-containing adaptors in TLR pathways
Mansell, A., Gay, N. & Kobe, B.
National Health and Medical Research Council (NHMRC) (Australia)
1/01/11 → 31/12/13
Project: Research