한빛사 논문
Abstract
Je-Kyung Ryu,1,2,9,10 Soo Jin Kim,3,9 Sang-Hyun Rah,1,2,7,8 Ji In Kang,3 Hi Eun Jung,3 Dongsun Lee,4 Heung Kyu Lee,3,4 Jie-Oh Lee,5 Beom Seok Park,6 Tae-Young Yoon,1,2,7,8,* and Ho Min Kim3,4,11,*
1National Creative Research Initiative Center for Single-Molecule Systems Biology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea
2Department of Physics, KAIST, Daejeon 34141, Korea
3Biomedical Science and Engineering Interdisciplinary Program, KAIST, Daejeon 34141, Korea
4Graduate School of Medical Science & Engineering, KAIST, Daejeon 34141, Korea
5Department of Chemistry, KAIST, Daejeon 34141, Korea
6Department of Biomedical Laboratory Science, College of Health Science, Eulji University, Seongnam 461-713, Korea
7Center for Nanomedicine, Institute for Basic Science (IBS), Yonsei University, Seoul 03722, Korea
8Yonsei-IBS Institute, Yonsei University, Seoul 03722, Korea
9Co-first author
10Present address: Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2629 HZ Delft, the Netherlands
11Lead Contact
*Correspondence : Tae-Young Yoon, Ho Min Kim
Summary
Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, binds Toll-like receptor 4 (TLR4)-MD2 complex and activates innate immune responses. LPS transfer to TLR4-MD2 is catalyzed by both LPS binding protein (LBP) and CD14. To define the sequential molecular interactions underlying this transfer, we reconstituted in vitro the entire LPS transfer process from LPS micelles to TLR4-MD2. Using electron microscopy and single-molecule approaches, we characterized the dynamic intermediate complexes for LPS transfer: LBP-LPS micelles, CD14-LBP-LPS micelle, and CD14-LPS-TLR4-MD2 complex. A single LBP molecule bound longitudinally to LPS micelles catalyzed multi-rounds of LPS transfer to CD14s that rapidly dissociated from LPB-LPS complex upon LPS transfer via electrostatic interactions. Subsequently, the single LPS molecule bound to CD14 was transferred to TLR4-MD2 in a TLR4-dependent manner. The definition of the structural determinants of the LPS transfer cascade to TLR4 may enable the development of targeted therapeutics for intervention in LPS-induced sepsis.
Keywords : innate immunity, LPS recognition, LPS transfer, LBP, CD14, TLR4/MD2, single-molecule fluorescence analysis, negative-stain EM
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