title: Molecular cloning and function characterization of a new macrophage-activating protein from Tremella fuciformis.
abstract: Silver ear mushroom ( Tremella fuciformis ) is an edible fungus with health benefits. In this study, we purified a new T. fuciformis protein (TFP) and demonstrated its ability to activate primary murine macrophages. The isolation procedure involved ammonium sulfate fractionation and ion exchange chromatography. TFP naturally formed a 24 kDa homodimeric protein and did not contain glycan residues. The TFP gene was cloned using the rapid amplification of cDNA ends method, and the cDNA sequence of TFP was composed of 408 nucleotides with a 336 nucleotide open reading frame encoding a 112 amino acid protein. TFP was capable of stimulating TNF-α, IL-1β, IL-1ra, and IL-12 production in addition to CD86/MHC class II expression, mRNA expression of M1-type chemokines, and nuclear NF-κB accumulation in murine peritoneal macrophage cells. Furthermore, TFP failed to stimulate TLR4-neutralized and TLR4-knockout macrophages, suggesting that TLR4 is a required receptor for TFP signaling on macrophages. Taken together, these results indicate that TFP may be an important bioactive compound from T. fuciformis that induces M1-polarized activation through a TLR4-dependent NF-κB signaling pathway.
authors.affiliation: Department of Horticulture and Landscape Architecture and ‡Center for Biotechnology, National Taiwan University , No. 1, Sec. 4, Roosevelt Road, Taipei 10673, Taiwan, R.O.C.
authors.forename: Chih-Liang
authors.initials: CL
authors.lastname: Hung
authors.forename: An-Ju
authors.initials: AJ
authors.lastname: Chang
authors.forename: Xhao-Kai
authors.initials: XK
authors.lastname: Kuo
authors.forename: Fuu
authors.initials: F
authors.lastname: Sheu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001487:Basidiomycota / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology*; D018664:Interleukin-12 / Q000235:genetics / Q000276:immunology; D053583:Interleukin-1beta / Q000235:genetics / Q000276:immunology; D008262:Macrophage Activation / Q000187:drug effects; D016215:Macrophage-Activating Factors / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology*; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology
chemical_list: D005656:Fungal Proteins; D053583:Interleukin-1beta; D016215:Macrophage-Activating Factors; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12