title: Resolving acetylated and phosphorylated proteins by neutral urea Triton-polyacrylamide gel electrophoresis: NUT-PAGE.
abstract: Protein acetylation and phosphorylation are key modifications that regulate both normal and pathological protein functions. The gel systems currently used for analyzing modified proteins require either expensive reagents or time-consuming second dimension electrophoresis. Here we present a neutral pH gel system that allows the analysis of acetylated and phosphorylated proteins. The neutral pH urea Triton-polyacrylamide gel electrophoresis (NUT-PAGE) system separates proteins based on their charge at pH 7.0 and generates discrete bands from each acetylated and/or phosphorylated species. In addition, the gel is composed of common and inexpensive laboratory reagents and requires only a single dimension of electrophoresis. We demonstrate the effectiveness of this system by analyzing the phosphorylated species of an acidic protein, α-synuclein, and both acetylated and phosphorylated species of a basic protein, histone H3. NUT-PAGE thus provides a cost-effective alternative for resolving acetylated and phosphorylated proteins, and potentially proteins with other post-translational modifications that alter net charge.
authors.affiliation: Cell and Molecular Biology Program, Michigan State University, East Lansing, MI.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Buehl
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI.
authors.forename: Xiexiong
authors.initials: X
authors.lastname: Deng
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI.
authors.forename: Mengyu
authors.initials: M
authors.lastname: Liu
authors.affiliation: Cell and Molecular Biology Program, Michigan State University, East Lansing, MI.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: McAndrew
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI.
authors.forename: Stacy
authors.initials: S
authors.lastname: Hovde
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI.
authors.forename: Xinjing
authors.initials: X
authors.lastname: Xu
authors.affiliation: Cell and Molecular Biology Program, Michigan State University, East Lansing, MI; Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI.
authors.forename: Min-Hao
authors.initials: MH
authors.lastname: Kuo
mesh_terms: D000107:Acetylation; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006367:HeLa Cells; D006657:Histones / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017830:Octoxynol / Q000737:chemistry; D010766:Phosphorylation / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D014508:Urea / Q000737:chemistry; D051844:alpha-Synuclein / Q000032:analysis* / Q000737:chemistry
chemical_list: D006657:Histones; D051844:alpha-Synuclein; D014508:Urea; D017830:Octoxynol
keywords: acetylation; histone H3; phosphorylation; α-synuclein
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