Functional glycoproteomics by integrated network assembly and partitioning
Abstract
The post-translational modification (PTM) of proteins by O-linked β-N-acetyl-D-glucosamine (O-GlcNAcylation) is widespread across the proteome during the lifespan of all multicellular organisms. However, nearly all functional studies have focused on individual protein modifications, overlooking the multitude of simultaneous O-GlcNAcylation events that work together to coordinate cellular activities. Here, we describeNetworking ofInteractors andSubstratEs (NISE), a novel, systems-level approach to rapidly and comprehensively monitor O-GlcNAcylation across the proteome. Our method integrates affinity purification-mass spectrometry (AP-MS) and site-specific chemoproteomic technologies with network generation and unsupervised partitioning to connect potential upstream regulators with downstream targets of O-GlcNAcylation. The resulting network provides a data-rich framework that reveals both conserved activities of O-GlcNAcylation such as epigenetic regulation as well as tissue-specific functions like synaptic morphology. Beyond O-GlcNAc, this holistic and unbiased systems-level approach provides a broadly applicable framework to study PTMs and discover their diverse roles in specific cell types and biological states.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jun 14, 2023
- Source ID
- 10.1101/2023.06.13.541482
Entities
People
- Andrew Schacht
- Annie Moradian
- Brett Lomenick
- Elizabeth H. Jensen
- John W Thompson
- Linda Hsieh-Wilson
- Matthew Griffin
- Michael Sweredoski
- Priya Choudhry
- Rita B. Aksenfeld
- S D Garbis
- Terry Kim
- Yao Xiao
- Yelena Koldobskaya