Resolving acetylated and phosphorylated proteins by neutral urea Triton-polyacrylamide gel electrophoresis: NUT-PAGE

BioTechniques

Christopher J. Buehl¹  Min-Hao Kuo¹  Xiexiong Deng²  Mengyu Liu²  Michael J McAndrew²  Stacy Hovde²  Xinjing Xu² 

[1] 1Cell and Molecular Biology Program, Michigan State University, East Lansing, MI;2Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI;

关键词: acetylation;    phosphorylation;    α-synuclein;    histone H3;   

DOI  :  10.2144/000114197

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown