Analysis of protein interaction complexes by advanced mass spectrometry workflows

The analysis of protein-protein interactions is critical for understanding cell growth control, and how aberrant connections contribute
to cancer and other diseases. Mass spectrometry is a critical tool in this field, but sample complexity, instrument dynamic range and
resolution limit some applications. This project will evaluate the use of the SelexION ion mobility device and High Resolution TOF
with Zeno pulsing for enhanced separation and detection of cross-linked peptides from protein complexes and to detect posttranslational
modifications of proteins, specifically phosphorylation. Ion mobility should separate phosphorylated peptides from one
another in a manner orthogonal to HPLC separation and increase sensitivity resulting in more comprehensive site analysis. Finally, a
critical component to understanding protein-protein interactions is the cellular response to drugs and/or mutations. For this,
quantitation is required, and we propose to investigate the use of a series of novel mass-defect reagents to allow for multiplexed DIA
quantitation of protein-protein interaction samples. By accessing High Resolution TOF with Zeno pulsing, the mass defect labels will
allow multiple samples to be analyzed at once, with quantitation resolved by mass measurement. This project will showcase new
instrumentation capabilities and provide feedback for instrument design and marketing material.

Faculty Supervisor:

Ann-Claude Gingras

Student:

Shen Zhang

Partner:

University of Toronto

Discipline:

Genetics

Sector:

Medical devices

University:

Program: