Profiling the specific reactivity of the proteome with non-directed activity-based probes

Gregory C. Adam, Benjamin F. Cravatt, Erik J. Sorensen

Research output: Contribution to journalArticle

125 Scopus citations

Abstract

Background: The field of proteomics aims to characterize dynamics in protein function on a global level. However, several classes of proteins, in particular low abundance proteins, remain difficult to characterize using standard proteomics technologies. Recently, chemical strategies have emerged that profile classes of proteins based on activity rather than quantity, thereby greatly facilitating the analysis of low abundance constituents of the proteome. Results: In order to expand the classes of proteins susceptible to analysis by activity-based methods, we have synthesized a library of biotinylated sulfonate esters and applied its members to complex proteomes under conditions that distinguish patterns of specific protein reactivity. Individual sulfonates exhibited unique profiles of proteome reactivity that in extreme cases appeared nearly orthogonal to one another. A robustly labeled protein was identified as a class I aldehyde dehydrogenase and shown to be irreversibly inhibited by members of the sulfonate library. Conclusions: Through screening the proteome with a non-directed library of chemical probes, diverse patterns of protein reactivity were uncovered. These probes labeled protein targets based on properties other than abundance, circumventing one of the major challenges facing contemporary proteomics research. Considering further that the probes were found to inhibit a target enzyme's catalytic activity, the methods described herein should facilitate the identification of compounds possessing both selective proteome reactivities and novel bioactivities.

Original languageEnglish (US)
Pages (from-to)81-95
Number of pages15
JournalChemistry and Biology
Volume8
Issue number1
DOIs
StatePublished - Jan 1 2001
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

Keywords

  • Activity-based
  • Aldehyde dehydrogenase
  • Probe
  • Proteomics
  • Sulfonate

Fingerprint Dive into the research topics of 'Profiling the specific reactivity of the proteome with non-directed activity-based probes'. Together they form a unique fingerprint.

  • Cite this