title: Scanning protein analysis of electrofocusing gels using X-ray fluorescence.
abstract: Recently, "metallomics," in addition to genomics and proteomics, has become a focus as a novel approach to identify sensitive fluctuations in homeostasis that accompany metabolic processes, such as stress responses, differentiation, and proliferation. Cellular elements and associated protein behavior provide important clues for understanding cellular and disease mechanism(s). It is important to develop a system for measuring the native status of the protein. In this study, we developed an original freeze-dried electrofocusing native gel over polyimide film (native-gel film) for scanning protein analysis using synchrotron radiation excited X-ray fluorescence (SPAX). To our knowledge, this is the first report detailing the successful mapping of metal-associated proteins of electrofocusing gels using X-ray fluorescence. SPAX can provide detection sensitivity equivalent to that of LA-ICP-MS. In addition to this increased sensitivity, SPAX has the potential to be combined with other X-ray spectroscopies. Our system is useful for further applications in proteomics investigating cellular element-associated protein behaviors and disease mechanisms.
authors.affiliation: Department of Precision Science and Technology, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Matsuyama
authors.forename: Akihiro
authors.initials: A
authors.lastname: Matsunaga
authors.forename: Shinichi
authors.initials: S
authors.lastname: Sakamoto
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Iida
authors.forename: Yoshinari
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Yukihito
authors.initials: Y
authors.lastname: Ishizaka
authors.forename: Kazuto
authors.initials: K
authors.lastname: Yamauchi
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Mari
authors.initials: M
authors.lastname: Shimura
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005453:Fluorescence; D007525:Isoelectric Focusing; D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D012015:Reference Standards; D013054:Spectrophotometry, Atomic; D013482:Superoxide Dismutase / Q000378:metabolism; D017356:Synchrotrons; D014965:X-Rays
chemical_list: D013482:Superoxide Dismutase