GST Pull-Down Assay to Measure Complex Formations

  • Sun-Yong Kim
  • Toshio HakoshimaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1893)


The GST pull-down assay is an intuitive and fast in vitro method for analyzing protein–protein or protein–ligand interactions and is comprised of a “bait” which is a GST-fused protein expressed in E. coli host or a baculovirus expression system and a “prey” which comprises putative binding partner protein(s) or other ligand molecule(s). This method is suitable for examining the direct interaction between two purified proteins and estimating the extent of the affinity.

Key words

GST pull-down assay Glutathione S-transferase Glutathione-conjugated resin Direct protein–protein interaction In vitro assay Screening of binding partners 


  1. 1.
    Fields S, Song OK (1989) A novel genetic system to detect protein–protein interactions. Nature 340:245–246CrossRefGoogle Scholar
  2. 2.
    Kim S-Y, Tachioka Y, Mori T, Hakoshima T (2016) Structural basis for autoinhibition and its relief of MOB1 in the Hippo pathway. Sci Rep 6:28488CrossRefGoogle Scholar
  3. 3.
    Kohler RS, Schmitz D, Cornils H, Hemmings BA, Hergovich A (2010) Differential NDR/LATS interactions with the human MOB family reveal a negative role for human MOB2 in the regulation of human NDR kinases. Mol Cell Biol 30:4507–4520CrossRefGoogle Scholar
  4. 4.
    Ni L, Zheng Y, Hara M, Pan D, Luo X (2015) Structural basis for Mob1-dependent activation of the core Mst-Lats kinase cascade in Hippo signaling. Genes Dev 29:1416–1431CrossRefGoogle Scholar
  5. 5.
    Smith DB, Johnson KS (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67:31–40CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Structural Biology LaboratoryNara Institute of Science and TechnologyNaraJapan

Personalised recommendations