Fig. 5
From: Structural and functional dissection of the DH and PH domains of oncogenic Bcr-Abl tyrosine kinase
Characterization of PIP binding to the PH domain. a Structure-based identification of the PIP-binding motif. The electrostatic surface representation indicates that the PIP-binding motif (black arrow) forms a positively charged patch on the PH surface. b The PIP-binding site is shown enlarged together with a D-myo-inositiol-4,5-bisphosphate (IP2) ligand fitted in the 2Fo–Fc electron-density map, contoured at 1.0 sigma. Critical residues in the binding motif are shown in sticks representation and colored by element. c PIP strips comparing the PIP-binding properties of PH wt, the PH R723A/K724A and R726A mutants, as well as the DH domain as a negative control. Individual signals on the strip were quantified relative to PH wild-type protein. d Binding of Bcr-Abl domains to surrogates of biological membranes in the liposome microarray (LiMA) assay. 0.1, 0.5, 1, and 2 μM of purified sfGFP-tagged Bcr-Abl DH, PH, DH–PH or Dbs DH–PH domains were incubated together with giant unilamellar liposomes containing the indicated concentrations of signaling lipid (in mol % of total lipids). PI3P, PI4P, PI5P, PI(3,4)P2, PI(3,5)P2, PI(4,5)P2, and PI(3,4,5)P3 are phosphoinositides (Supplementary Data 2, 3). PC phosphatidylcholine, PA phosphatidic acid, PE phosphatidylethanolamine, PI phosphatidylinositol PS phosphatidylserine, LPA lyso PA, LPC lyso PC, S1P sphingosine-1-phosphate, NBI normalized binding intensity. Not determined values are indicated with a star (*). Values are means (n = 3)
