title: Inhibition of clathrin/dynamin-dependent internalization interferes with LPS-mediated TRAM-TRIF-dependent signaling pathway.
abstract: Recognition of lipopolysaccharide (LPS) by Toll-like receptor 4 (TLR4) activates two district proinflammatory signaling pathway and initiates LPS internalization. To investigate roles of LPS internalization, a traditionally regarded metabolic pathway for LPS, in regulation of these two pathways, three internalization inhibitors, monodansylcadaverine (MDC, a clathrin inhibitor), dynasore (DS, a dynamin inhibitor) and chloroquine (CQ, an endosome acidifying maturation inhibitor) were applied to induce internalization dysfunction in macrophages. Results showed MDC and DS affected LPS internalization but did not interfere with their colocalization. Additionally, they decreased cytokines and chemokines release and inhibited signaling molecules activation mediated by TRAM-TRIF-dependent pathway as determined by protein array. In contrast, CQ did not inhibit LPS internalization but affected the colocalization. It also suppressed macrophage activation mediated by both MyD88-dependent and TRAM-TRIF-dependent pathways. The above data indicated that LPS internalization was clathrin/dynamin dependent and it was essential for activation of TRAM-TRIF-dependent signaling pathway.
authors.affiliation: Medical Research Center, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Wang
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Hongwei
authors.initials: H
authors.lastname: Cao
authors.forename: Yongling
authors.initials: Y
authors.lastname: Lu
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.forename: Jiang
authors.initials: J
authors.lastname: Zheng
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D018925:Chemokines / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D002966:Clathrin / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D034281:Dynamins / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D006835:Hydrazones / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000276:immunology* / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D018123:Receptors, Interleukin / Q000378:metabolism*; D015398:Signal Transduction; D051197:Toll-Like Receptor 4 / Q000276:immunology / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D002103:Cadaverine; D018925:Chemokines; D002738:Chloroquine; D002966:Clathrin; D016207:Cytokines; D034281:Dynamins; D006835:Hydrazones; D008070:Lipopolysaccharides; D053594:Myeloid Differentiation Factor 88; D018123:Receptors, Interleukin; D051197:Toll-Like Receptor 4; C507780:Myd88 protein, mouse; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; C476286:Ticam1 protein, mouse; C476416:Ticam2 protein, mouse; C493487:Tlr4 protein, mouse; C008542:monodansylcadaverine