The first Ras-binding domain of RGS14 interacts with Ras and Rap family GTPases in vitro.
figureposted on 2009-03-25, 01:03 authored by Francis S. Willard, Melinda D. Willard, Adam J. Kimple, Meera Soundararajan, Emily A. Oestreich, Xiaoyan Li, Nathaniel A. Sowa, Randall J. Kimple, Declan A. Doyle, Channing J. Der, Mark J. Zylka, William D. Snider, David P. Siderovski
GST-fusion proteins of indicated full-length or truncated RGS14 were incubated with lysates from HEK293T cells transfected with wild-type (WT) or mutationally-activated (GV) HA-tagged H-Ras (A), Rap2A (B), or FLAG-tagged Rap2B (C). Protein complexes were precipitated with glutathione agarose, washed, and resolved by SDS-PAGE and immunoblot (IB) (bottom panels). Experimental samples were also analyzed by immunoblot to ensure equivalent loading of GTPases (middle panels). Precipitation of GST-fusion proteins was confirmed by SDS-PAGE and Coomassie Blue staining (top panels). Data are representative of 3 or more independent experiments.