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. 2009:1:29-41.
doi: 10.1002/9780470559277.ch090138.

Activity-Based Protein Profiling (ABPP) and Click Chemistry (CC)-ABPP by MudPIT Mass Spectrometry

Anna E Speers  1 , Benjamin F Cravatt
Affiliations

Activity-Based Protein Profiling (ABPP) and Click Chemistry (CC)-ABPP by MudPIT Mass Spectrometry

Anna E Speers et al. Curr Protoc Chem Biol. 2009.

Abstract

Activity-based protein profiling (ABPP) is a chemical proteomic method for functional interrogation of enzymes within complex proteomes. This Unit presents a protocol for in vitro and in vivo labeling using ABPP and Click Chemistry (CC)-ABPP probes for in-depth profiling using the Multi-dimensional Protein Identification Technology (MudPIT) analysis platform.

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Figures

Figure 1
Figure 1
The ABPP-MudPIT method for high-content proteomic analysis of enzyme activities. A proteome is labeled with a biotinylated ABPP probe, and labeled proteins are affinity enriched on streptavidin beads. After on-bead trypin digest, the tryptic peptides are analyzed by MudPIT for identification and quantification (e.g., via spectra counting). Box: The probe labeled peptides can also be eluted from the streptavidin beads for MS analysis of labeling sites.
See this image and copyright information in PMC

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