RAP-MS: A Method to Identify Proteins that Interact Directly with a Specific RNA Molecule in Cells

Methods Mol Biol. 2018:1649:473-488. doi: 10.1007/978-1-4939-7213-5_31.

Abstract

RNA molecules interact with proteins to perform a variety of functions in living cells. The binding partners of many RNAs, in particular the newly discovered class of long noncoding RNAs (lncRNAs), remain largely unknown. RNA antisense purification coupled with mass spectrometry (RAP-MS) is a method that enables the identification of direct and specific protein interaction partners of a specific RNA molecule. Because RAP-MS uses direct RNA-protein cross-linking methods coupled along with highly denaturing purification conditions, RAP-MS provides a short list of high confidence protein interactors.

Keywords: Antisense nucleic acid capture; Mass spectrometry; Protein purification; RNA purification; RNA-binding proteins; RNA–protein interactions; UV cross-linking.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Fractionation
  • Detergents
  • Isotope Labeling
  • Mass Spectrometry / methods*
  • Mice
  • Mouse Embryonic Stem Cells / metabolism*
  • Peptides / isolation & purification
  • Protein Binding
  • RNA, Antisense / metabolism*
  • RNA-Binding Proteins / metabolism*

Substances

  • Detergents
  • Peptides
  • RNA, Antisense
  • RNA-Binding Proteins