Proteomic analysis of microtubule-associated proteins during macrophage activation.
Classical activation of macrophages induces a wide range of signaling and vesicle trafficking events to produce a more aggressive cellular phenotype. The microtubule (MT) cytoskeleton is crucial for the regulation of immune responses. In the current study, we used a large scale proteomics approach to analyze the change in protein composition of the MT-associated protein (MAP) network by macrophage stimulation with the inflammatory cytokine interferon-gamma and the endotoxin lipopolysaccharide. Overall the analysis identified 409 proteins that bound directly or indirectly to MTs. Of these, 52 were up-regulated 2-fold or greater and 42 were down-regulated 2-fold or greater after interferon-gamma/lipopolysaccharide stimulation. Bioinformatics analysis based on publicly available binary protein interaction data produced a putative interaction network of MAPs in activated macrophages. We confirmed the up-regulation of several MAPs by immunoblotting and immunofluorescence analysis. More detailed analysis of one up-regulated protein revealed a role for HSP90beta in stabilization of the MT cytoskeleton during macrophage activation.
Patel
Prerna C
PC
Department of Biological Sciences, University of Toronto Scarborough, 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
Fisher
Katherine H
KH
Yang
Eric C C
EC
Deane
Charlotte M
CM
Harrison
Rene E
RE
D000818:Animals; D019295:Computational Biology / Q000379:methods; D003599:Cytoskeleton / Q000378:metabolism; D030562:Databases, Protein; D005189:False Positive Reactions; D005786:Gene Expression Regulation; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism; D008262:Macrophage Activation; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008954:Models, Biological; D040901:Proteomics / Q000379:methods*
D008070:Lipopolysaccharides; D008869:Microtubule-Associated Proteins