title: A novel ML protein from Manduca sexta may function as a key accessory protein for lipopolysaccharide signaling.
abstract: Lipopolysaccharide (LPS) present on the outer membrane of Gram-negative bacteria is one of the most important pathogen-associated molecular patterns and a potent elicitor in innate immunity. In human, TLR4 (Toll-like receptor 4) and MD-2 (myeloid differiation-2) form a receptor complex to transduce the LPS signal into cells. However, in invertebrates, receptors that recognize LPS have not been determined. Here we report the purification, characterization and cDNA cloning of an ML (MD-2-related lipid-recognition) protein from the tobacco hornworm Manduca sexta. The full-length cDNA of this M. sexta ML protein, named MsML-1, is 532bp with an open reading frame of 456bp that encodes a polypeptide of 151 amino acids containing an ML domain. MsML-1 is a secreted glycoprotein and its mRNA is expressed in fat body and hemocytes. The expression level of MsML-1 mRNA in fat body and hemocytes as well as MsML-1 protein in hemolymph are not induced by immune challenge. Recombinant MsML-1 protein specifically binds to LPS from several Gram-negative bacteria and LPS Re mutant, as well as to lipid A, but not to KDO (2-keto-3-deoxyoctonate). Our results suggest that MsML-1 may function as a key accessory protein for LPS signaling in M. sexta against Gram-negative bacterial infection.
authors.affiliation: Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri-Kansas City, Kansas City, MO 64110, USA.
authors.forename: Jing-qun
authors.initials: JQ
authors.lastname: Ao
authors.forename: Erjun
authors.initials: E
authors.lastname: Ling
authors.forename: Xiang-jun
authors.initials: XJ
authors.lastname: Rao
authors.forename: Xiao-Qiang
authors.initials: XQ
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005216:Fat Body / Q000276:immunology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006434:Hemocytes / Q000276:immunology; D006458:Hemolymph / Q000276:immunology; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008050:Lipid A / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology* / Q000494:pharmacology; D018402:Manduca / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology; D016415:Sequence Alignment; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D008050:Lipid A; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D011994:Recombinant Proteins