TLR4/MD-2 activation by a synthetic agonist with no similarity to LPS

Ying Wang; Lijing Su; Matthew D. Morin; Brian T. Jones; Landon R. Whitby; Murali M.R.P. Surakattula; Hua Huang; Hexin Shi; Jin Huk Choi; Kuan Wen Wang; Eva Marie Y. Moresco; Michael Berger; Xiaoming Zhan; Hong Zhang; Dale L. Boger; Bruce Beutler

doi:10.1073/pnas.1525639113

TLR4/MD-2 activation by a synthetic agonist with no similarity to LPS

Ying Wang, Lijing Su, Matthew D. Morin, Brian T. Jones, Landon R. Whitby, Murali M.R.P. Surakattula, Hua Huang, Hexin Shi, Jin Huk Choi, Kuan Wen Wang, Eva Marie Y. Moresco, Michael Berger, Xiaoming Zhan, Hong Zhang, Dale L. Boger, Bruce Beutler^*

^*Corresponding author for this work

Department of Immunology and Cancer Research

Research output: Contribution to journal › Article › peer-review

111 Scopus citations

Abstract

Structurally disparate molecules reportedly engage and activate Toll-like receptor (TLR) 4 and other TLRs, yet the interactions that mediate binding and activation by dissimilar ligands remain unknown. We describe Neoseptins, chemically synthesized peptidomimetics that bear no structural similarity to the established TLR4 ligand, lipopolysaccharide (LPS), but productively engage the mouse TLR4 (mTLR4)/ myeloid differentiation factor 2 (MD-2) complex. Neoseptin-3 activates mTLR4/MD-2 independently of CD14 and triggers canonical myeloid differentiation primary response gene 88 (MyD88)- and Toll-interleukin 1 receptor (TIR) domain-containing adaptor inducing IFN-beta (TRIF)- dependent signaling. The crystal structure mTLR4/MD-2/Neoseptin-3 at 2.57-Å resolution reveals that Neoseptin-3 binds as an asymmetrical dimer within the hydrophobic pocket of MD-2, inducing an active receptor complex similar to that induced by lipid A. However, Neoseptin-3 and lipid A form dissimilar molecular contacts to achieve receptor activation; hence strong TLR4/MD-2 agonists need not mimic LPS.

Original language	American English
Pages (from-to)	E884-E893
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	113
Issue number	7
DOIs	https://doi.org/10.1073/pnas.1525639113
State	Published - 16 Feb 2016

Bibliographical note

Funding Information:
We thank Ian Wilson for the initial design of the mouse TLR4VLR and MD2-protein A hybrid constructs for coexpression of the complex; Jose Rizo-Rey for help with the NMR experiments; Diana Tomchick and Srinivasan Raghunathan for help with X-ray data collection and structure determination; and Peter Jurek and Anne Murray for assistance with manuscript preparation. Results shown in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center at Advanced Photon Source. Argonne is operated by UChicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research under Contract DE-AC02-06CH11357. This work was supported by NIH/National Institute of General Medical Sciences (NIGMS) Grant R01GM104496 (to H.Z.) and NIH/National Institute of Allergy and Infectious Diseases (NIAID) Grants U24 AI082657 (to B.B. and D.L.B.) and U19 AI100627 (to B.B.).

Keywords

Crystal structure
Innate immunity
Neoseptins
Peptidomimetic compounds
Proinflammatory response

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10.1073/pnas.1525639113

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Wang, Y., Su, L., Morin, M. D., Jones, B. T., Whitby, L. R., Surakattula, M. M. R. P., Huang, H., Shi, H., Choi, J. H., Wang, K. W., Moresco, E. M. Y., Berger, M., Zhan, X., Zhang, H., Boger, D. L., & Beutler, B. (2016). TLR4/MD-2 activation by a synthetic agonist with no similarity to LPS. Proceedings of the National Academy of Sciences of the United States of America, 113(7), E884-E893. https://doi.org/10.1073/pnas.1525639113

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abstract = "Structurally disparate molecules reportedly engage and activate Toll-like receptor (TLR) 4 and other TLRs, yet the interactions that mediate binding and activation by dissimilar ligands remain unknown. We describe Neoseptins, chemically synthesized peptidomimetics that bear no structural similarity to the established TLR4 ligand, lipopolysaccharide (LPS), but productively engage the mouse TLR4 (mTLR4)/ myeloid differentiation factor 2 (MD-2) complex. Neoseptin-3 activates mTLR4/MD-2 independently of CD14 and triggers canonical myeloid differentiation primary response gene 88 (MyD88)- and Toll-interleukin 1 receptor (TIR) domain-containing adaptor inducing IFN-beta (TRIF)- dependent signaling. The crystal structure mTLR4/MD-2/Neoseptin-3 at 2.57-{\AA} resolution reveals that Neoseptin-3 binds as an asymmetrical dimer within the hydrophobic pocket of MD-2, inducing an active receptor complex similar to that induced by lipid A. However, Neoseptin-3 and lipid A form dissimilar molecular contacts to achieve receptor activation; hence strong TLR4/MD-2 agonists need not mimic LPS.",

keywords = "Crystal structure, Innate immunity, Neoseptins, Peptidomimetic compounds, Proinflammatory response",

author = "Ying Wang and Lijing Su and Morin, {Matthew D.} and Jones, {Brian T.} and Whitby, {Landon R.} and Surakattula, {Murali M.R.P.} and Hua Huang and Hexin Shi and Choi, {Jin Huk} and Wang, {Kuan Wen} and Moresco, {Eva Marie Y.} and Michael Berger and Xiaoming Zhan and Hong Zhang and Boger, {Dale L.} and Bruce Beutler",

note = "Funding Information: We thank Ian Wilson for the initial design of the mouse TLR4VLR and MD2-protein A hybrid constructs for coexpression of the complex; Jose Rizo-Rey for help with the NMR experiments; Diana Tomchick and Srinivasan Raghunathan for help with X-ray data collection and structure determination; and Peter Jurek and Anne Murray for assistance with manuscript preparation. Results shown in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center at Advanced Photon Source. Argonne is operated by UChicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research under Contract DE-AC02-06CH11357. This work was supported by NIH/National Institute of General Medical Sciences (NIGMS) Grant R01GM104496 (to H.Z.) and NIH/National Institute of Allergy and Infectious Diseases (NIAID) Grants U24 AI082657 (to B.B. and D.L.B.) and U19 AI100627 (to B.B.).",

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Wang, Y, Su, L, Morin, MD, Jones, BT, Whitby, LR, Surakattula, MMRP, Huang, H, Shi, H, Choi, JH, Wang, KW, Moresco, EMY, Berger, M, Zhan, X, Zhang, H, Boger, DL & Beutler, B 2016, 'TLR4/MD-2 activation by a synthetic agonist with no similarity to LPS', Proceedings of the National Academy of Sciences of the United States of America, vol. 113, no. 7, pp. E884-E893. https://doi.org/10.1073/pnas.1525639113

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AU - Su, Lijing

AU - Morin, Matthew D.

AU - Jones, Brian T.

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AU - Surakattula, Murali M.R.P.

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AU - Shi, Hexin

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AU - Wang, Kuan Wen

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AU - Berger, Michael

AU - Zhan, Xiaoming

AU - Zhang, Hong

AU - Boger, Dale L.

AU - Beutler, Bruce

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AB - Structurally disparate molecules reportedly engage and activate Toll-like receptor (TLR) 4 and other TLRs, yet the interactions that mediate binding and activation by dissimilar ligands remain unknown. We describe Neoseptins, chemically synthesized peptidomimetics that bear no structural similarity to the established TLR4 ligand, lipopolysaccharide (LPS), but productively engage the mouse TLR4 (mTLR4)/ myeloid differentiation factor 2 (MD-2) complex. Neoseptin-3 activates mTLR4/MD-2 independently of CD14 and triggers canonical myeloid differentiation primary response gene 88 (MyD88)- and Toll-interleukin 1 receptor (TIR) domain-containing adaptor inducing IFN-beta (TRIF)- dependent signaling. The crystal structure mTLR4/MD-2/Neoseptin-3 at 2.57-Å resolution reveals that Neoseptin-3 binds as an asymmetrical dimer within the hydrophobic pocket of MD-2, inducing an active receptor complex similar to that induced by lipid A. However, Neoseptin-3 and lipid A form dissimilar molecular contacts to achieve receptor activation; hence strong TLR4/MD-2 agonists need not mimic LPS.

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TLR4/MD-2 activation by a synthetic agonist with no similarity to LPS

Abstract

Bibliographical note

Keywords

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