Sunday, March 4, 2012, 1:00 – 4:00 pm. Tuition is $100. 

Organizer:  Ileana M. Cristea (Princeton University)

Course Description

Dynamic protein interactions carry out the majority of the processes within a cell, including cellular responses to environmental stimuli and pathogens. Isolation of protein complexes and characterization of protein-protein interactions provide critical insights into their biological functions. An ideal isolation would maintain the protein-protein interaction or the protein assembly as close as possible to the original state in the cell. Therefore, proteomic-based methodologies that can access stable and transient interactions are invaluable for diverse studies, such as those of cell cycle or pathogen infection that require characterization of temporal and spatial protein interactions. This course will cover fundamental and practical aspects of studying protein interactions. Topics discussed will include: 1) protein function considerations for workflow design, 2) cell lysis methods for efficient protein extraction, 3) critical choices for optimizing an immunoaffinity purification experiment, including resin type and speed of isolation, 4) denaturing and non-denaturing methods of eluting captured protein complexes, 5) assessing the specificity of interactions using bioinformatics approaches, metabolic labeling with stable isotopes, or peptide labeling with isobaric tags, 6) challenges for assessing direct or indirect interactions, and 7) aspects of data analysis and generation of interaction networks. As we will gradually cover fundamental and more advanced topics concerning protein interactions, the course will be appropriate for both beginner- and advanced-level participants. Detailed protocols will be provided, and enough time will be set aside for discussing these topics from either a mass spectrometry or biology perspective. 

Level: Beginner to advanced (the course will gradually cover fundamental to advanced topics concerning protein interactions).

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