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J. Biol. Chem., Vol. 276, Issue 24, 21129-21135, June 15, 2001
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From The Scripps Research Institute, The structural features of some proteins of the
innate immune system involved in mediating responses to microbial
pathogens are highly conserved throughout evolution. Examples include
members of the Drosophila Toll (dToll) and the mammalian
Toll-like receptor (TLR) protein families. Activation of
Drosophila Toll is believed to occur via an endogenous
peptide rather than through direct binding of microbial products to the
Toll protein. In mammals there is a growing consensus that
lipopolysaccharide (LPS) initiates its biological activities through a
heteromeric receptor complex containing CD14, TLR4, and at least one
other protein, MD-2. LPS binds directly to CD14 but whether LPS then
binds to TLR4 and/or MD-2 is not known. We have used transient
transfection to express human TLRs, MD-2, or CD14 alone or in different
combinations in HEK 293 cells. Interactions between LPS and these
proteins were studied using a chemically modified, radioiodinated LPS
containing a covalently linked, UV light-activated cross-linking group
(125I-ASD-Re595 LPS). Here we show that LPS is cross-linked
specifically to TLR4 and MD-2 only when co-expressed with CD14. These
data support the contention that LPS is in close proximity to the three known proteins of its membrane receptor complex. Thus, LPS binds directly to each of the members of the tripartite LPS receptor complex.
Lipopolysaccharide Is in Close Proximity to Each of the Proteins
in Its Membrane Receptor Complex
TRANSFER FROM CD14 TO TLR4 AND MD-2*
,,, and
Department of
Immunology and the § Department of Neuropharmacology,
La Jolla, California 92037
*
This work was supported in part by National Institutes of
Health Grants AI15136 (to R. J. U.), GM28485 (to R. J. U.), GM37696 (to R. J. U.), AI32021 (to P. S. T.), and HL23584 (to P. S. T.). This is Publication
136-IMM from The Scripps Research Institute, Department of Immunology.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.:
858-784-8219; Fax: 858-784-8239; E-mail: ulevitch@scripps.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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G. Meng, A. Grabiec, M. Vallon, B. Ebe, S. Hampel, W. Bessler, H. Wagner, and C. J. Kirschning Cellular Recognition of Tri-/Di-palmitoylated Peptides Is Independent from a Domain Encompassing the N-terminal Seven Leucine-rich Repeat (LRR)/LRR-like Motifs of TLR2 J. Biol. Chem., October 10, 2003; 278(41): 39822 - 39829. [Abstract] [Full Text] [PDF]
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 S. Akashi, S.-i. Saitoh, Y. Wakabayashi, T. Kikuchi, N. Takamura, Y. Nagai, Y. Kusumoto, K. Fukase, S. Kusumoto, Y. Adachi, et al. Lipopolysaccharide Interaction with Cell Surface Toll-like Receptor 4-MD-2: Higher Affinity than That with MD-2 or CD14 J. Exp. Med., October 6, 2003; 198(7): 1035 - 1042. [Abstract] [Full Text] [PDF]
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M. A. Freudenberg, C. Kalis, Y. Chvatchko, T. Merlin, M. Gumenscheimer, and C. Galanos Role of interferons in LPS hypersensitivity Innate Immunity, October 1, 2003; 9(5): 308 - 312. [Abstract] [PDF]
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B. Beutler, K. Hoebe, X. Du, and R. J. Ulevitch How we detect microbes and respond to them: the Toll-like receptors and their transducers J. Leukoc. Biol., October 1, 2003; 74(4): 479 - 485. [Abstract] [Full Text] [PDF]
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J. Y. Lee, J. Ye, Z. Gao, H. S. Youn, W. H. Lee, L. Zhao, N. Sizemore, and D. H. Hwang Reciprocal Modulation of Toll-like Receptor-4 Signaling Pathways Involving MyD88 and Phosphatidylinositol 3-Kinase/AKT by Saturated and Polyunsaturated Fatty Acids J. Biol. Chem., September 26, 2003; 278(39): 37041 - 37051. [Abstract] [Full Text] [PDF]
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K. Sau, S. S. Mambula, E. Latz, P. Henneke, D. T. Golenbock, and S. M. Levitz The Antifungal Drug Amphotericin B Promotes Inflammatory Cytokine Release by a Toll-like Receptor- and CD14-dependent Mechanism J. Biol. Chem., September 26, 2003; 278(39): 37561 - 37568. [Abstract] [Full Text] [PDF]
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H. Shimomura, M. Matsuura, S. Saito, Y. Hirai, Y. Isshiki, and K. Kawahara Unusual Interaction of a Lipopolysaccharide Isolated from Burkholderia cepacia with Polymyxin B Infect. Immun., September 1, 2003; 71(9): 5225 - 5230. [Abstract] [Full Text] [PDF]
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 K. Yamazaki, K. Ueki-Maruyama, T. Oda, K. Tabeta, Y. Shimada, H. Tai, T. Nakajima, H. Yoshie, D. Herawati, and G.J. Seymour Single-nucleotide Polymorphism in the CD14 Promoter and Periodontal Disease Expression in a Japanese Population Journal of Dental Research, August 1, 2003; 82(8): 612 - 616. [Abstract] [Full Text] [PDF]
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N. Qureshi, P.-Y. Perera, J. Shen, G. Zhang, A. Lenschat, G. Splitter, D. C. Morrison, and S. N. Vogel The Proteasome as a Lipopolysaccharide-Binding Protein in Macrophages: Differential Effects of Proteasome Inhibition on Lipopolysaccharide-Induced Signaling Events J. Immunol., August 1, 2003; 171(3): 1515 - 1525. [Abstract] [Full Text] [PDF]
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P. A. Thompson, P. S. Tobias, S. Viriyakosol, T. N. Kirkland, and R. L. Kitchens Lipopolysaccharide (LPS)-binding Protein Inhibits Responses to Cell-bound LPS J. Biol. Chem., August 1, 2003; 278(31): 28367 - 28371. [Abstract] [Full Text] [PDF]
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R. Dziarski, K. A. Platt, E. Gelius, H. Steiner, and D. Gupta Defect in neutrophil killing and increased susceptibility to infection with nonpathogenic gram-positive bacteria in peptidoglycan recognition protein-S (PGRP-S)-deficient mice Blood, July 15, 2003; 102(2): 689 - 697. [Abstract] [Full Text] [PDF]
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 E. S. Van Amersfoort, T. J. C. Van Berkel, and J. Kuiper Receptors, Mediators, and Mechanisms Involved in Bacterial Sepsis and Septic Shock Clin. Microbiol. Rev., July 1, 2003; 16(3): 379 - 414. [Abstract] [Full Text] [PDF]
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M. Sato, H. Sano, D. Iwaki, K. Kudo, M. Konishi, H. Takahashi, T. Takahashi, H. Imaizumi, Y. Asai, and Y. Kuroki Direct Binding of Toll-Like Receptor 2 to Zymosan, and Zymosan-Induced NF-{kappa}B Activation and TNF-{alpha} Secretion Are Down-Regulated by Lung Collectin Surfactant Protein A J. Immunol., July 1, 2003; 171(1): 417 - 425. [Abstract] [Full Text] [PDF]
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 K. A. Walton, A. L. Cole, M. Yeh, G. Subbanagounder, S. R. Krutzik, R. L. Modlin, R. M. Lucas, J. Nakai, E. J. Smart, D. K. Vora, et al. Specific Phospholipid Oxidation Products Inhibit Ligand Activation of Toll-Like Receptors 4 and 2 Arterioscler. Thromb. Vasc. Biol., July 1, 2003; 23(7): 1197 - 1203. [Abstract] [Full Text] [PDF]
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 I. Ben-Shlomo, S. Yu Hsu, R. Rauch, H. W. Kowalski, and A. J. W. Hsueh Signaling Receptome: A Genomic and Evolutionary Perspective of Plasma Membrane Receptors Involved in Signal Transduction Sci. Signal., June 17, 2003; 2003(187): re9 - re9. [Abstract] [Full Text] [PDF]
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F. Bihl, L. Salez, M. Beaubier, D. Torres, L. Lariviere, L. Laroche, A. Benedetto, D. Martel, J.-M. Lapointe, B. Ryffel, et al. Overexpression of Toll-Like Receptor 4 Amplifies the Host Response to Lipopolysaccharide and Provides a Survival Advantage in Transgenic Mice J. Immunol., June 15, 2003; 170(12): 6141 - 6150. [Abstract] [Full Text] [PDF]
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T. Ohnishi, M. Muroi, and K.-i. Tanamoto MD-2 Is Necessary for the Toll-Like Receptor 4 Protein To Undergo Glycosylation Essential for Its Translocation to the Cell Surface Clin. Vaccine Immunol., May 1, 2003; 10(3): 405 - 410. [Abstract] [Full Text] [PDF]
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C. F. Ortega-Cava, S. Ishihara, M. A. K. Rumi, K. Kawashima, N. Ishimura, H. Kazumori, J. Udagawa, Y. Kadowaki, and Y. Kinoshita Strategic Compartmentalization of Toll-Like Receptor 4 in the Mouse Gut J. Immunol., April 15, 2003; 170(8): 3977 - 3985. [Abstract] [Full Text] [PDF]
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G. E. D. Mullen, M. N. Kennedy, A. Visintin, A. Mazzoni, C. A. Leifer, D. R. Davies, and D. M. Segal The role of disulfide bonds in the assembly and function of MD-2 PNAS, April 1, 2003; 100(7): 3919 - 3924. [Abstract] [Full Text] [PDF]
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R. Malley, P. Henneke, S. C. Morse, M. J. Cieslewicz, M. Lipsitch, C. M. Thompson, E. Kurt-Jones, J. C. Paton, M. R. Wessels, and D. T. Golenbock Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection PNAS, February 18, 2003; 100(4): 1966 - 1971. [Abstract] [Full Text] [PDF]
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 H-J Yoon, J H Shin, S H Yang, D-W Chae, H Kim, D-S Lee, H L Kim, S Kim, J S Lee, and Y S Kim Association of the CD14 gene -159C polymorphism with progression of IgA nephropathy J. Med. Genet., February 1, 2003; 40(2): 104 - 108. [Abstract] [Full Text] [PDF]
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K. Kawasaki, H. Nogawa, and M. Nishijima Identification of Mouse MD-2 Residues Important for Forming the Cell Surface TLR4-MD-2 Complex Recognized by Anti-TLR4-MD-2 Antibodies, and for Conferring LPS and Taxol Responsiveness on Mouse TLR4 by Alanine-Scanning Mutagenesis J. Immunol., January 1, 2003; 170(1): 413 - 420. [Abstract] [Full Text] [PDF]
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E. Latz, A. Visintin, E. Lien, K. A. Fitzgerald, B. G. Monks, E. A. Kurt-Jones, D. T. Golenbock, and T. Espevik Lipopolysaccharide Rapidly Traffics to and from the Golgi Apparatus with the Toll-like Receptor 4-MD-2-CD14 Complex in a Process That Is Distinct from the Initiation of Signal Transduction J. Biol. Chem., November 27, 2002; 277(49): 47834 - 47843. [Abstract] [Full Text] [PDF]
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 P. Knuefermann, S. Nemoto, A. Misra, N. Nozaki, G. Defreitas, S. M. Goyert, B. A. Carabello, D. L. Mann, and J. G. Vallejo CD14-Deficient Mice Are Protected Against Lipopolysaccharide-Induced Cardiac Inflammation and Left Ventricular Dysfunction Circulation, November 12, 2002; 106(20): 2608 - 2615. [Abstract] [Full Text] [PDF]
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 A. Biragyn, P. A. Ruffini, C. A. Leifer, E. Klyushnenkova, A. Shakhov, O. Chertov, A. K. Shirakawa, J. M. Farber, D. M. Segal, J. J. Oppenheim, et al. Toll-Like Receptor 4-Dependent Activation of Dendritic Cells by beta -Defensin 2 Science, November 1, 2002; 298(5595): 1025 - 1029. [Abstract] [Full Text] [PDF]
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M. Muroi, T. Ohnishi, and K.-i. Tanamoto Regions of the Mouse CD14 Molecule Required for Toll-like Receptor 2- and 4-mediated Activation of NF-kappa B J. Biol. Chem., October 25, 2002; 277(44): 42372 - 42379. [Abstract] [Full Text] [PDF]
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X. Ye and S. F. Liu Lipopolysaccharide Down-regulates Sp1 Binding Activity by Promoting Sp1 Protein Dephosphorylation and Degradation J. Biol. Chem., August 23, 2002; 277(35): 31863 - 31870. [Abstract] [Full Text] [PDF]
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 M.M. Monick and G.W. Hunninghake Activation of second messenger pathways in alveolar macrophages by endotoxin Eur. Respir. J., July 1, 2002; 20(1): 210 - 222. [Abstract] [Full Text] [PDF]
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S. Liu, D. J. Gallo, A. M. Green, D. L. Williams, X. Gong, R. A. Shapiro, A. A. Gambotto, E. L. Humphris, Y. Vodovotz, and T. R. Billiar Role of Toll-Like Receptors in Changes in Gene Expression and NF-{kappa}B Activation in Mouse Hepatocytes Stimulated with Lipopolysaccharide Infect. Immun., July 1, 2002; 70(7): 3433 - 3442. [Abstract] [Full Text] [PDF]
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D. Iwaki, H. Mitsuzawa, S. Murakami, H. Sano, M. Konishi, T. Akino, and Y. Kuroki The Extracellular Toll-like Receptor 2 Domain Directly Binds Peptidoglycan Derived from Staphylococcus aureus J. Biol. Chem., June 28, 2002; 277(27): 24315 - 24320. [Abstract] [Full Text] [PDF]
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F. Re and J. L. Strominger Monomeric Recombinant MD-2 Binds Toll-like Receptor 4 Tightly and Confers Lipopolysaccharide Responsiveness J. Biol. Chem., June 21, 2002; 277(26): 23427 - 23432. [Abstract] [Full Text] [PDF]
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M. T. Abreu, E. T. Arnold, L. S. Thomas, R. Gonsky, Y. Zhou, B. Hu, and M. Arditi TLR4 and MD-2 Expression Is Regulated by Immune-mediated Signals in Human Intestinal Epithelial Cells J. Biol. Chem., May 31, 2002; 277(23): 20431 - 20437. [Abstract] [Full Text] [PDF]
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J. Fan, A. Kapus, P. A. Marsden, Y. H. Li, G. Oreopoulos, J. C. Marshall, S. Frantz, R. A. Kelly, R. Medzhitov, and O. D. Rotstein Regulation of Toll-Like Receptor 4 Expression in the Lung Following Hemorrhagic Shock and Lipopolysaccharide J. Immunol., May 15, 2002; 168(10): 5252 - 5259. [Abstract] [Full Text] [PDF]
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S. Li, V. M. Holers, S. A. Boackle, and C. M. Blatteis Modulation of Mouse Endotoxic Fever by Complement Infect. Immun., May 1, 2002; 70(5): 2519 - 2525. [Abstract] [Full Text] [PDF]
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 N. Tuaillon, D. F. Shen, R. B. Berger, B. Lu, B. J. Rollins, and C.-C. Chan MCP-1 Expression in Endotoxin-Induced Uveitis Invest. Ophthalmol. Vis. Sci., May 1, 2002; 43(5): 1493 - 1498. [Abstract] [Full Text] [PDF]
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