Titanium Dioxide Photocatalytic Polymerization of Acrylamide for Gel Electrophoresis (TIPPAGE) of Proteins and Structural Identification by Mass Spectrometry

• Published in: Scientific reports 2016-02, Vol.6 (1), p.20981-20981, Article 20981
• Main Authors: Zhang, Wenyang, Yuan, Zhiwei, Huang, Lulu, Kang, Jie, Jiang, Ruowei, Zhong, Hongying
• Format: Article
• Language: English
• Subjects: 639/638/11/296; 639/638/11/872; 82/29; Acrylamide; Acrylamide - chemistry; Acrylamide - radiation effects; Ammonium; Ammonium Sulfate - chemistry; Animals; Biological samples; Caseins - isolation & purification; Cell lines; Degassing; Electrophoresis; Electrophoresis, Polyacrylamide Gel - methods; Ethylenediamines - chemistry; Free radicals; Gel electrophoresis; Gels; Histones; Histones - isolation & purification; Humanities and Social Sciences; Hydroxyl radicals; Liver - chemistry; Mass Spectrometry; Mass spectroscopy; multidisciplinary; Nanoparticles; Octoxynol - chemistry; Phosphorylation; Photocatalysis; Photochemical Processes; Polymerization; Proteins; Pyronine; Rats; Reagents; Science; Scientific imaging; Titanium; Titanium - chemistry; Titanium dioxide; Ultraviolet radiation; Ultraviolet Rays; Urea; Urea - chemistry
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep20981

Polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry has been well established for separating, identifying and quantifying protein mixtures from cell lines, tissues or other biological samples. The copolymerization process of acrylamide and bis-acrylamide is the key to mastering this powerful technique. In general, this is a vinyl addition reaction initiated by free radical-generating reagents such as ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) under basic pH and degassing experimental condition. We report herein a photocatalytic polymerization approach that is based on photo-generated hydroxyl radicals with nanoparticles of titanium dioxide. It was shown that the polymerization process is greatly accelerated in acidic condition when ultraviolet light shots on the gel solution containing TiO 2 nanoparticles without degassing. This feature makes it very useful in preparing Triton X-100 acid urea (TAU) gel that has been developed for separating basic proteins such as histones and variants in acidic experimental condition. Additionally, the presence of titanium dioxide in the gel not only improves mechanistic property of gels but also changes the migration pattern of different proteins that have different affinities to titanium dioxide.