Chem 219: Mass Spectrometry-based Proteomics

Spring 2024

Proteomics is broadly concerned with gaining new knowledge on global protein composition at the cellular, sub-cellular, or protein complex level and how their functions are modulated by posttranslational dynamics, e.g. kinome or ubiquitylome/proteasome homeostasis; or the effects of molecular defects on aberrant signaling pathways and networks, etc. Mass spectrometry is the technology of choice for detecting this information due to its ability to analyze complex samples for both discovery and quantification.

Course Description: This course will focus on the practical aspects (i.e. experimental and mass spectral interpretation) involved in the identification of proteins and their covalent modifications. It will cover the fundamental principles of currently important mass spectrometry instrument platforms. It will provide an overview of key scientific problems that are being tackled and solved at the protein-level relevant to cell function/dysfunction; the detection and assignment of protein posttranslational modifications; and studies of site-specific dynamics/relative quantitation. It will also cover studies on the architecture of protein complexes and machines.

Simultaneously, dry laboratory work will be performed to learn how to carry out the interpretation of complex mixtures of peptides isolated from cells involved in current active research projects. A block of time will be scheduled for class discussion of the experimental results you obtain.

Lecture Schedule: April 1 – 19, 2024:
Tu and Th 1-3pm, Mission Bay Campus BH-413

Lecture Lecturer Topic
Tu 2nd ALB Fundamentals: Ionization, Instrumentation; ion optics, resolution and mass accuracy; why these are important at protein vs peptide level.
Th 4th RJC Protein Identification. Basics of peptide fragmentation processes. Database searching. How to measure the reliability of assignments.
Tu 9th JO Sample preparation: Gels and Chromatography; IP/Affinity Tags, Digestion. What shouldn't be in the sample - Contaminants.
Th 11th AU Large scale quantitation strategies (Label-free, SILAC, TMT, PRM, DIA).
Tu 16th JM Posttranslational modifications: Protein vs peptide analysis. PTM enrichment, PTM cross-talk.
Th 18th MT Architecture of protein complexes and machines (chemical cross-linking).

If you want to participate please email Al Burlingame, so that instructors can have an idea of the number of people who will attend.

Lecturers

ALB - Al Burlingame
RJC - Robert Chalkley
JM - Jason Maynard
JO - Juan Oses
MT - Mike Trnka
AU - Anatoly Urisman

National Institute of General Medical SciencesAdelson Medical Research Foundation