FIGURE 8.

Effect of mutated PLD2 on physiological assays. A, shown is the effect of representative mutants (R172C, ΔIle-255–Val-267 and ΔIle-306–Ala-310) on GTP binding of Rac2 over time. B, shown is the effect of PX substitution mutants and PH deletion mutants on PLD2 lipase activity. Phagocytosis and chemotaxis activities of PLD2 mutants (mut) are shown in panels C and D, respectively. The PH point mutations were made in pcDNA3.1-mycPLD2 (full-length). Cells were transfected with a constant amount of PLD2 DNA (2 μg) and used for either phagocytosis of zymosan particles or chemotaxis using Transwell inserts. Data are the means ± S.E. from at least three independent experiments performed in duplicate. * or # represent differences between means, as determined by ANOVA, above or below the negative control levels. PLD2 residues involved in chemotaxis and phagocytosis in vivo are outlined in a gray box. E, shown are immunofluorescence photomicrographs of cells overexpressing PLD2-WT and mutants during phagocytosis of zymosan beads.