Skip to main content
. 2012 Oct 5;287(49):41268–41276. doi: 10.1074/jbc.M112.419127

FIGURE 3.

FIGURE 3.

Amino acids responsible for lipid-binding specificity of the FYVE domain. A, amino acid sequence of the protrudin-FYVE domain was compared with that of the EEA1-FYVE domain. Lipid-binding pocket (purple) of the EEA1-FYVE domain is composed of a WXXD motif, R + HHC + XCG, and RVC regions. Corresponding sites in the protrudin-FYVE domain are colored pink. The CXXC motif or the predicted amino acids in the protrudin-FYVE domain were distinctly marked as green and red. B, a model of PtdIns(4,5)P2 binding to the protrudin-FYVE domain. The PtdIns(4,5)P2 molecule is shown in stick, and the FYVE domain in an electrostatic surface model. The positively charged residues (blue) presumed to interact with PtdIns(4,5)P2 are labeled. The structure is shown using PyMOL software (DeLano Scientific LLC). C, images of cells expressing the wild type FYVE domain, K367A/R369A, R386A, and 4A. Bars, 10 μm. D, quantitative analysis of the expression level of the FYVE domain on the PM (WT, n = 10; K367A, n = 28; K367A/R369A, n = 11; R381A, n = 20; R386A, n = 19; 2A, n = 12; 4A, n = 10). Error bars, mean ± S.E. E, images of cells expressing the full-length of protrudin (WT), protrudin containing K367A/R369A and 4A (K367A/R369A/R381A/K386A) mutations, and the FYVE domain-deleted protrudin (ΔFYVE), respectively. Bars, 10 μm.