Crystallization and X-ray diffraction analysis of the N-terminal domain of the Toll-like receptor signalling adaptor protein TRIF/TICAM-1

Ullah, M. Obayed, Ve, Thomas, Dkhar, Jameris, Alaidarous, Mohammed, Ericsson, Daniel J., Sweet, Matthew J., Mansell, Ashley and Kobe, Bostjan (2013) Crystallization and X-ray diffraction analysis of the N-terminal domain of the Toll-like receptor signalling adaptor protein TRIF/TICAM-1. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 69 7: 766-770. doi:10.1107/S174430911301419X

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Author Ullah, M. Obayed
Ve, Thomas
Dkhar, Jameris
Alaidarous, Mohammed
Ericsson, Daniel J.
Sweet, Matthew J.
Mansell, Ashley
Kobe, Bostjan
Title Crystallization and X-ray diffraction analysis of the N-terminal domain of the Toll-like receptor signalling adaptor protein TRIF/TICAM-1
Journal name Acta Crystallographica Section F: Structural Biology and Crystallization Communications   Check publisher's open access policy
ISSN 1744-3091
Publication date 2013-06-01
Year available 2013
Sub-type Article (original research)
DOI 10.1107/S174430911301419X
Open Access Status File (Publisher version)
Volume 69
Issue 7
Start page 766
End page 770
Total pages 5
Place of publication Malden, MA, United States
Publisher Wiley-Blackwell
Collection year 2014
Language eng
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-NTDA66M/L113M are reported. Thin plate-like crystals of native TRIF-NTD obtained using polyethylene glycol 3350 as precipitant diffracted X-rays to 1.9 Å 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 Å resolution. The TRIF-NTD and TRIF-NTDA66M/L113M crystals have the symmetry of space groups P2 12121 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.
Keyword Adaptor proteins
Innate immunity
Introduction of additional methionine residues
Selenomethionine labelling
Toll-like receptors
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
School of Chemistry and Molecular Biosciences
Institute for Molecular Bioscience - Publications
 
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Created: Fri, 12 Jul 2013, 23:38:52 EST by Mrs Louise Nimwegen on behalf of School of Chemistry & Molecular Biosciences