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Volumn 309, Issue 5734, 2005, Pages 581-585

Structural biology: Crystal structure of human toll-like receptor 3 (TLR3) ectodomain

Author keywords

[No Author keywords available]

Indexed keywords

CRYSTAL STRUCTURE; HYDROGEN BONDS; OLIGOMERS; RNA; SOLENOIDS;

EID: 23044445303     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.1115253     Document Type: Article
Times cited : (520)

References (51)
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    • note
    • 6-tag was cloned into pAcGP67A vector. The protein was expressed with HiS cells and a baculovirus expression system. We infected the cells with baculovirus using a multiplicity of infection of 3, and protein was harvested from the media after 4 days. We purified the protein using Ni-NTA, ion-exchange (MonoQ, Pharmacia), and size-exclusion chromatography (Superdex 200, Pharmacia). The purified protein was concentrated to 15 mg/ml in 10 mM TrisHCl at pH 7.5, 100 mM NaCl, 2 mM EDTA, and 5 mM DTT.
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    • note
    • 2- molecules. Data collection and refinement statistics are summarized in table 51.
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    • note
    • Ribonuclease inhibitor was the first LRR structure and is the most similar to TLR3, in that it forms an almost complete horseshoe but with different dimensions and a different number of repeats.
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    • note
    • The point of insertion at position 11 was determined by comparison with neighboring LRR motif structures. It is often referred to as an insertion at position 15 from previous sequence comparisons.
  • 22
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    • note
    • The first 12 residues of the C-cap region corresponding to the consensus LRR motif of TLR3 form a modified, incomplete LRR structure.
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    • note
    • The sugars are modeled here as oligomannans, which predominate in insect cells as in our construct, with up to nine mannoses and two N- acetylglucosamines.
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    • note
    • The poly(I:C) was purchased from InvivoGen, San Diego. For experimental details, see fig. 57.
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    • note
    • We thank R. Ulevitch for kindly providing template TLR3 DNA; X. Dai and R. Stanfield for help with data collection; staff members at the Stanford Synchrotron Radiation Laboratory beamline 11-1 and Advanced Light Source 8.2.1; and B. Beutler and R. Ulevitch for helpful comments and suggestions. Supported by NIH grant nos. AI-42266 and CA-58896 (I.A.W.); a Skaggs Institute post-doctoral fellowship (J.C.); and NIH postdoctoral training grant no. T32 AI077606, a TSRI Skaggs predoctoral fellowship, and a TSRI Jairo H. Arévelo fellowship (M.S.K.). This is manuscript number 17439-MB from the Scripps Research Institute. Structure factors and the coordinates are deposited in the Protein Data Bank under accession code 1ZIW.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.