title: Phosphoproteome analysis of Drosophila melanogaster embryos.
abstract: Protein phosphorylation is a key regulatory event in most cellular processes and development. Mass spectrometry-based proteomics provides a framework for the large-scale identification and characterization of phosphorylation sites. Here, we used a well-established phosphopeptide enrichment and identification strategy including the combination of strong cation exchange chromatography, immobilized metal affinity chromatography, and high-accuracy mass spectrometry instrumentation to study phosphorylation in developing Drosophila embryos. In total, 13,720 different phosphorylation sites were discovered from 2702 proteins with an estimated false-discovery rate (FDR) of 0.63% at the peptide level. Because of the large size of the data set, both novel and known phosphorylation motifs were extracted using the Motif-X algorithm, including those representative of potential ordered phosphorylation events.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhai
authors.forename: Judit
authors.initials: J
authors.lastname: Villén
authors.forename: Sean A
authors.initials: SA
authors.lastname: Beausoleil
authors.forename: Julian
authors.initials: J
authors.lastname: Mintseris
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D029721:Drosophila Proteins / Q000032:analysis* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D008954:Models, Biological; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction / Q000502:physiology; D053719:Tandem Mass Spectrometry / Q000379:methods; D014357:Trypsin / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D010750:Phosphoproteins; D014357:Trypsin