Professor Jennifer S. Brodbelt of the University of Texas at Austin is supported by the Chemical Measurement and Imaging Program in the Division of Chemistry to develop analytical methodologies based on mass spectrometry for the analysis of intact proteins. These experiments provide information on the locations of multiple posttranslational modifications, which makes it possible to study the influence of those groups on protein function and hence allow better understanding of the structure and properties of biological systems. The proposed research promises to overcome some of the technical hurdles associated with the activation and dissociation of intact proteins to generate meaningful fragment ion maps that can be used for protein identification. If successful, the proposed work will positively contribute to the Human Proteome Project, an on-going effort to catalog all human proteins. It will also facilitate the use of top-down protein mass spectrometry for solving problems in the areas of biotherapeutics, structural biology, and mechanistic biochemistry. In this context, the project will provide new avenues for the identification of proteins and hence advance our understanding of life processes.
In this project, studies are being conducted to develop ultraviolet photodissociation (UVPD) as a versatile strategy for the analysis of proteins by mass spectrometry, and to explore the mechanistic, structural, and experimental features that play a role in the ultraviolet photodissociation process. The project has five separate but related specific aims: (1) to explore the parameters for effective UVPD of intact proteins; (2) to develop a hybrid electron transfer dissociation- ultraviolet photodissociation (ETD/UVPD) methods for mass spectrometric fragmentation of intact proteins; (3) to modify the charge state of proteins for more effective ultraviolet photodissociation; (4) to modulate the selectivity of UVPD by the attachment of specific chromophores to the proteins; and (5) to develop UVPD methodologies for structural biology applications with a focus on determining the structures of intact proteins and the binding interactions of native protein complexes.
During the course of this project, students will be trained in advanced mass spectrometry, laser optics, derivatization reactions, protein processing, and bioinformatics, and summer boot camps will be set up to increase awareness of the potential applications of advanced mass spectrometry for solving biological problems.