title: Proteomic analysis of the systemic immune response of Drosophila.
abstract: Improvements in two-dimensional gel electrophoresis, mass spectrometry, and bioinformatics provide new tools to characterize proteins involved in a physiological process, such as the immune response of the insect model Drosophila melanogaster. Profiling of the proteins present in the hemolymph (insect blood) of noninfected flies versus flies infected with bacteria or fungi was performed by two-dimensional gel electrophoresis, silver or Coomassie staining, and image analysis. Through this differential analysis, more than 70 out of 160 spots were up- or down-regulated by at least 5-fold after microbial infection. Coomassie staining, in-gel digestion, and database searches yielded the identity of a series of proteins that are directly involved in the Drosophila immune system. This included proteases, protease inhibitors, and recognition molecules such as prophenoloxydase-activating enzymes, serpins, and Gram-negative binding protein-like. Proteins with a potential function in the immune response were also identified, such as an odorant binding protein, peptidylglycine alpha-hydroxylating monooxygenase, and transferrin, affording new candidates for further investigation of innate immune mechanisms. Moreover, several molecules resulting from the cleavage of proteins were detected after the fungal infection. Altogether, this first differential proteomic analysis of the immune response of Drosophila paves the way for the study of proteins affected during innate immunity.
authors.affiliation: Institut de Biologie Moléculaire et Cellulaire, 15 rue René Descartes, 67084 Strasbourg Cedex, France.
authors.forename: Francine
authors.initials: F
authors.lastname: Levy
authors.forename: Philippe
authors.initials: P
authors.lastname: Bulet
authors.forename: Laurence
authors.initials: L
authors.lastname: Ehret-Sabatier
mesh_terms: D000818:Animals; D001424:Bacterial Infections / Q000276:immunology; D029721:Drosophila Proteins / Q000276:immunology / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000378:metabolism; D005658:Fungi / Q000378:metabolism; D006458:Hemolymph / Q000276:immunology / Q000378:metabolism*; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D016982:Micrococcus luteus / Q000378:metabolism; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D029721:Drosophila Proteins; D000949:Histocompatibility Antigens Class II; D020543:Proteome