Fig. 3
From: Structural basis of Gip1 for cytosolic sequestration of G protein in wide-range chemotaxis
Recognition of the prenyl group on Gγ by the hydrophobic cavity of Gip1. a Schematic diagram of Gγ modification. C-terminal cysteine (C66) is geranylgeranylated and methylated (Me). b Subcellular localization of Gγ in a living cell. Flag-Flag-GFP (F2G) tag alone (vector) or F2G-tagged Gγ(WT) or Gγ(ΔCAAX) in gγΔ cells. Scale bar, 5 μm. c Co-immunoprecipitation of Gγ or Gip1. F2G-Gγ(WT) or F2G-Gγ(ΔCAAX) was expressed in gγΔ cells (left). GFP-Flag-tagged Gip1 (Gip1-GFPF) was coexpressed with Gγ(WT) or Gγ(ΔCAAX) in gγΔ cells (right). Pull-down samples of Gγ (left) and Gip1 (right) were immunoblotted with the indicated antibodies. d In vitro interaction between Gβγ and purified Gip1. Gβγ subunits were bound to beads and incubated with purified full-length Gip1. e Competitive dissociation of G proteins from Gip1 by geranylgeranyl pyrophosphate (GG-pyroP). The data were normalized relative to the band intensities without GG-pyroP and presented as the mean ± SD of three independent experiments (n = 3, *P < 0.05, ***P < 0.001 versus 0% GG-pyroP, two-tailed unpaired Student’s t-test)
