Global characterization of RNA modifying enzymes with RNA-mediated activity-based protein profiling (RNABPP)

Nathan J. Yu, Qais Z. Jaber, Ralph E. Kleiner

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Post-transcriptional RNA modifications can regulate RNA function and play an important role in gene expression. Studying RNA modifying enzymes and their associated modifications remains a considerable challenge. Here we describe the RNA-mediated activity-based protein profiling (RNABPP) methodology, a chemoproteomic strategy for profiling the activity of RNA modifying enzymes in their native context. RNABPP relies upon metabolic RNA labeling with modified ribonucleoside-based probes, combined with protein-RNA enrichment and quantitative proteomics. The RNABPP approach is a general strategy based on chemical reactivity and enzyme mechanism, making it suitable for probing multiple classes of RNA modifying enzymes across diverse biological systems.

Original languageEnglish (US)
Title of host publicationChemical and Genomic Methods in Nucleic Acid Biology
EditorsBrandt F. Eichman
PublisherAcademic Press Inc.
Pages111-125
Number of pages15
ISBN (Print)9780443297243
DOIs
StatePublished - Jan 2024

Publication series

NameMethods in Enzymology
Volume705
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Keywords

  • activity-based protein profiling
  • epitranscriptome
  • modified nucleosides
  • RNA modifications
  • RNABPP

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