Journal: PLoS ONE

Article Title: Autoinducers Act as Biological Timers in Vibrio harveyi

doi: 10.1371/journal.pone.0048310

Figure Lengend Snippet: Inverted membrane vesicles prepared from E. coli TKR2000 containing full-length LuxQ were incubated with purified LuxP, purified LuxU and (where indicated) with 10 μM AI-2 (A). The phosphorylation reaction was started by adding 100 μM [γ- 32 P] ATP at time 0. At the indicated times, the reaction was terminated, and radiolabeled proteins were separated by SDS-PAGE, and visualized by autoradiography. The arrow indicates phosphorylated LuxU. Phosphorylated LuxU was quantified with ImageQuant using [γ- 32 P] ATP as standard (B). Phosphorylation experiments were also performed in the presence or absence of 10 μM HAI-1 using membrane vesicles containing full-length LuxN and phosphorylated LuxU was quantified accordingly (B).

Article Snippet: Different dilutions of [γ- 32 P] ATP were used to generate a calibration curve for quantification of the signal intensities of phosphorylated proteins using ImageQuant software (Molecular Dynamics V5.0; GE Healthcare).

Techniques: Membrane, Incubation, Purification, Phospho-proteomics, SDS Page, Autoradiography