Fig. 5
From: Structural basis of Gip1 for cytosolic sequestration of G protein in wide-range chemotaxis
Tryptophan mutagenesis to induce steric blockade in the cavity. a Representation of tryptophan-mutated residues. Mutated residues are shown as stick models. Magenta and cyan colours show the residues that lead to cytosolic Gα reduced and the residues with no effects, respectively. The surface of the cavity is coloured dark grey. b Subcellular localization of Gα2 and Gγ labelled with TMR and Gip1-GFPF in a living cell in the presence of LatA. Representative images of the top 25% of severely impaired mutants are shown. Scale bar, 5 μm. c Co-immunoprecipitation of Gip1 with Gβγ. The same mutants shown in b were analysed. The data were normalized relative to the band intensities of wild-type Gip1 and are presented as the mean ± SD of at least three independent experiments (n = 5 (Vector, WT), 4 (I306W, I166W, V190W), and 3 (L300W, L211W), ***P < 0.001 versus wild-type, two-tailed unpaired Student’s t-test)
