Toward a global characterization of the phosphoproteome in prostate cancer cells: identification of phosphoproteins in the LNCaP cell line.
Protein phosphorylation plays a major role in most cell-signaling pathways in all eukaryotic cells. Disruptions in phosphorylation-mediated cell-signaling events are associated with various diseases, including cancer. Here, we applied a fully non-gel-based methodology to obtain an initial panel of phosphoproteins from the LNCaP human prostate cancer cell line. The analytical strategy involved enrichment of phosphopeptides by immobilized metal ion affinity chromatography, the use of POROS Oligo R3 to capture phosphopeptides that were not retained with a C18 packing, and gas-phase fractionation in the m/z dimension to extend the dynamic range of the LC-MS/MS analysis. In this pilot investigation, 137 phosphorylation sites in 81 phosphoproteins were identified. The characterized phosphoproteins include kinases, co-regulators of steroid receptors, and a number of cancer-related proteins.
Giorgianni
Francesco
F
Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee Health Science Center, Memphis, TN, USA.
Zhao
Yingxin
Y
Desiderio
Dominic M
DM
Beranova-Giorgianni
Sarka
S
D000595:Amino Acid Sequence; D002457:Cell Extracts / Q000032:analysis / Q000737:chemistry*; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D030562:Databases, Protein; D006801:Humans; D008297:Male; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D010865:Pilot Projects; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D015398:Signal Transduction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D002457:Cell Extracts; D010750:Phosphoproteins; D020543:Proteome