601 Development of a comprehensive two-dimensional gel database of rat liver proteins useful for toxicological hazard identification

Abstract

After promising approaches during the past decade, the development of publicly available databases for rat liver proteins has been slowed down. Emerging proteomic technologies still depend on data generated by 2D-electrophoresis (2-DE), so we decided to develop a new 2-DE map for male Wistar rat liver proteins in the context of a molecular toxicological joint research project. 2-DE separation was performed using the common IPG technique developed by Goerg and coworkers. Spots were excised from 2D gels using a spot picker. After in-gel digestion with trypsin, proteins were identified by using MALDI-MS (MALDI-TOF) and LC-MS/MS (ESI-Quad-TOF). Available protein information, i.e. protein ID, molecular weight, isoelectrical point and EC-number were combined into an HTML document, which lead to a clickable HTML protein map. Over 300 proteins were identified, among those 140 enzymes and 100 structural proteins. This map could be the basis for a publicly accessible HTML document and could provide quick information to identify protein expression patterns of toxicological relevance. Among the 300 identified proteins we found a plenty of common toxicological marker proteins. Patterns of protein expression may contribute mechanistic data to assess toxic and carcinogenic effects of chemicals.