title: Protein phosphorylation in irradiated human melanoma cells.
abstract: In the present study, we examined the response of confluent, primary human fibroblasts and cells of a melanoma (YUSAC2) cell line to ionizing radiation mediated through post-translational protein phosphorylation. Since the purpose of our study was to identify novel radiation-induced phosphoproteins in the DNA damage stress response of melanoma cells, we were primarily interested in changes in protein phosphoserine expression at early times after irradiation. Our rationale was that by examining the overall protein phosphorylation profile (the phosphoproteome) in irradiated cells, we might discover novel radiation-induced phosphoproteins that distinguish fibroblasts from melanoma cells. Cell proteins were separated by gel electrophoresis and phosphoproteins were identified by Western blot analysis using nonspecific anti-phosphoamino acid antibodies. This approach was not pursued previously since adequate antibodies for examining global protein phosphoserine expression were unavailable. While some radiation-induced phosphoprotein changes in high-abundance proteins were identified, in general the sensitivity of this approach was not sufficient to detect changes in low-abundance, regulatory proteins. Characterization of these phosphoproteins will require greater enrichment of low-abundance proteins.
authors.affiliation: Department of Radiation Oncology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA. ray.warters@hsc.utah.edu
authors.forename: Raymond L
authors.initials: RL
authors.lastname: Warters
authors.forename: Dustin L
authors.initials: DL
authors.lastname: Williams
authors.forename: Sergey B
authors.initials: SB
authors.lastname: Zhuplatov
authors.forename: Chris D
authors.initials: CD
authors.lastname: Pond
authors.forename: Sancy A
authors.initials: SA
authors.lastname: Leachman
mesh_terms: D045744:Cell Line, Tumor; D004307:Dose-Response Relationship, Radiation; D015972:Gene Expression Regulation, Neoplastic / Q000528:radiation effects*; D006801:Humans; D008545:Melanoma / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation / Q000528:radiation effects; D011829:Radiation Dosage
chemical_list: D009363:Neoplasm Proteins; D010750:Phosphoproteins