Dr. Michael Goshe
Office Location: Polk Hall 43
MS Laboratory Supervisor |
Mr. Kevin Blackburn
Office Location: Polk Hall 49
Lab Location: Polk Hall 42
Lab Phone: 919-515-0831
Biological Mass Spectrometry Overview
Mass spectrometry is a powerful technique used frequently for both qualitative and quantitative characterization of proteins and other biomolecules as well as their interactions. In a typical MS experiment, molecules are ionized, accelerated into a mass analyzer, separated on the basis of mass-to-charge ratio (m/z), and detected. In another type of experiment known as MS/MS, one or more "precursor" ions are fragmented inside a collision cell in the mass spectrometer, and the resulting "product" ions mass analyzed. The resulting product ion spectrum may be used to provide insights into molecular structure based on the pattern of product ions generated. In combination with other techniques such as liquid chromatography, gel electrophoresis, chemical crosslinking, and co-immunoprecipitation, a variety of complex biological problems may be addressed.
The mass spectrometry facility in Molecular and Structural Biochemistry is used to support a variety of research efforts through collaboration with Dr. Goshe. Although not a fee-for-service core facility, a large number of collaborative efforts are supported for faculty across departments and colleges, including generation of preliminary data in support of planned grant submissions on a case by case basis. Ongoing efforts can be broadly categorized as follows.
Proteomics: Characterization of the composition and dynamics of proteomes in plant and animal systems using a variety of approaches including label-free quantitation, SILIP, SILAC, and other custom labeling techniques. Determination of protein interaction partners through the use of co-immunoprecipitaton and other affinity based techniques.
Characterization of Post-Translational Modifications: Site assignment and quantitative characterization of post-translational modifications including phosphorylation, ubiquitination, and others. Site-specific stoichiometry measurements in response to various stimuli or environmental conditions may be determined.
Protein Structure: Through the use of a variety of novel chemical crosslinking reagents, protein-protein interactions are studied. Insights into protein structure and folding may be obtained through the analysis of individual proteins following chemical crosslinking. Similarly, intramolecular disulfide bonding patterns within proteins may be determined by MS. In addition, quantitative compositional analysis of protein macromolecular complexes often provides insights into the structure and function of the complex or individual components.