PROTOMAP

PROTOMAP maps in vivo substrates of proteolytic events by combining one-dimensional SDS-PAGE with mass spectrometry to provide proteome-wide topography and magnitude of proteolytic fragments.

Key Features:

• 1D SDS-PAGE and mass spectrometry integration: Integrates one-dimensional SDS-PAGE and mass spectrometry to detect gel migration shifts and map proteolytic fragment topography.
• Proteome-wide coverage: Enables global mapping of proteolytic events across the entire proteome.
• Caspase substrate identification: Applied to the intrinsic apoptotic pathway to identify 91 characterized caspase substrates and 170 previously unreported cleavage targets.
• Fragment topography and magnitude visualization: Provides direct visualization of the topography and magnitude of proteolytic fragments, revealing persistent fragments corresponding to discrete protein domains.

Scientific Applications:

• Apoptosis research: Generates proteome-wide maps of proteolytic events induced by the intrinsic apoptotic pathway.
• Proteolytic pathway characterization: Characterizes proteolytic pathways to study protein activation, translocation, and degradation.
• Effector fragment analysis: Identifies persistent domain-sized fragments to study generation of active effector proteins resulting from proteolysis.
• Translational research: Informs therapeutic strategies targeting dysregulated proteolytic pathways by revealing substrate specificity and cleavage patterns.

Methodology:

Integration of one-dimensional SDS-PAGE and mass spectrometry to detect shifts in gel migration patterns and map the topography of proteolytic fragments across the proteome.