title: Biochemical and functional analysis of TIR domain containing protein from Brucella melitensis.
abstract: Toll/interleukin-1 like receptors are evolutionarily conserved proteins in eukaryotes that play crucial role in pathogen recognition and innate immune responses. Brucella are facultative intracellular bacterial pathogens causing brucellosis in animal and human hosts. Brucella behave as a stealthy pathogen by evading the immune recognition or suppressing the TLR signaling cascades. Brucella encode a TIR domain containing protein, TcpB, which suppresses NF-kappaB activation as well as pro-inflammatory cytokine secretion mediated by TLR2 and TLR4 receptors. TcpB targets the TIRAP mediated pathway to suppress TLR signaling. With the objective of detailed characterization, we have over expressed and purified TcpB from Brucella melitensis in native condition. The purified protein exhibited lipid-binding properties and cell permeability. NF-kappaB inhibition property of endogenous TcpB has also been demonstrated. The data provide insight into the mechanism of action of TcpB in the intracellular niche of Brucella.
authors.affiliation: Department of Pathobiological Sciences, University of Wisconsin-Madison, WI, USA.
authors.forename: Girish K
authors.initials: GK
authors.lastname: Radhakrishnan
authors.forename: Gary A
authors.initials: GA
authors.lastname: Splitter
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017347:Brucella melitensis / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D058619:Maltose-Binding Proteins; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D033902:Periplasmic Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D037521:Virulence Factors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D058619:Maltose-Binding Proteins; D016328:NF-kappa B; D033902:Periplasmic Binding Proteins; D010716:Phosphatidylinositols; D011993:Recombinant Fusion Proteins; C587630:TcpB protein, Brucella melitensis; D037521:Virulence Factors
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