Professor Nicolas C. Polfer of the University of Florida is supported by the Chemical Measurement and Imaging (CMI) Program in the Division of Chemistry to study the chemistry that takes place when proteins are analyzed by mass spectrometry. The project will expand the scope and improve the fidelity of mass spectrometric characterization of the fragmentation patterns of peptide ions generated from parent proteins. This, in turn, improves the reliability of the high throughput identifications of proteins in complex biological samples. Advances in life sciences depend on the integration of reliable protein identification results with complimentary genomic and metabolomic information. In this context, the project will provide new avenues for the identification of proteins and hence advance our understanding of life processes. In addition, the spectroscopic techniques developed in this work may also lead to applications in other areas of chemistry, such as mechanistic organic chemistry.
The analytical mass spectrometric and spectroscopic methods the research team is developing are focused on probing the chemistry that takes place when peptides are fragmented by collision-induced dissociation (CID) for the purpose of peptide and protein sequencing by mass spectrometry. Analytical laser spectroscopy techniques, in particular infrared multiphoton dissociation (IRMPD) and two-laser pump-probe schemes in a cryogenically-cooled 3-D ion trap, will be employed to: 1) provide a deeper understanding of labile and radical rearrangement chemistries that take place, 2) study the dynamics of these rearrangement reactions, and 3) develop methodologies that enable more control in the dissociation chemistry, for improved structural determination of peptides and proteins. The proposed research is of significance to de novo protein sequencing and for the location of posttranslational modifications on the peptide sequence.
In the course of this research, graduate students are being trained in state-of-the-art mass spectrometric and spectroscopic techniques. The technologies and the results of the proposed research will be communicated to high school students through workshops aimed at high school science teachers.
