Figure 3.
Specific binding of VfPIP210–312 to Vfphot1a. A, Binding was determined in vitro by protein-blot analysis using GST-VfPIP210–312 as a probe (lanes 1–4) for various phototropins and was detected with anti-GST antibody. His6-tagged- Vfphot1a and Vfphot1b were expressed in E. coli, and His6-tagged Atphot1 was expressed in insect cells. All these recombinant proteins were subjected to SDS-PAGE and blotted onto nitrocellulose membranes. As a control, analysis was done for His6-tagged Vfphot1a using GST alone (lane 5). VfPIP indicates V. faba phototropin 1a interacting protein. B, Western analyses for various phototropin species were done to confirm the presence of these phototropins using the respective antibodies (lanes 1–3, anti-Vfphot antibodies; lane 4, anti-Atphot1 antibodies).