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Single-step affinity purification of ERK signaling complexes using the streptavidin-binding peptide (SBP) tag

  • University of Massachusetts Boston

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Elucidation of biological functions of signaling proteins is facilitated by studying their protein–protein interaction networks. Affinity purification combined with mass spectrometry (AP-MS) has become a favorite method to study protein complexes. Here we describe a procedure for single-step purification of ERK (Rolled) and associated proteins from Drosophila cultured cells. The use of the streptavidin-binding peptide (SBP) tag allows for a highly efficient isolation of native ERK signaling complexes, which are suitable for subsequent analysis by mass spectrometry. Our analysis of the ERK interactome has identified both known and novel signaling components. This method can be easily adapted for SBP-based purification of protein complexes in any expression system.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages113-126
Number of pages14
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1487
ISSN (Print)1064-3745

ASJC Scopus Subject Areas

  • Molecular Biology
  • Genetics

Keywords

  • Affinity purification
  • Drosophila
  • ERK
  • Mass spectrometry
  • SBP
  • Streptavidin-binding peptide

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