Figure 2

Extreme termini of Gα ₁₆ are not required for TPR1 interaction. (A) Schematic representation of the C25, C44, and N30 chimeras. Predicted secondary structures are illustrated as boxes (α helices) or ovals (β strands) above the chimeras. Closed areas represent human Gα₁₆ sequence while those in open shapes signify the corresponding sequence of human Gα_z. (B-D) HEK 293 cells were transiently co-transfected with FLAG-TPR1 and the wild-type or constitutively active mutants of Gα₁₆, Gα_z, C25, C44, N30, or pcDNA1. Cell lysates from the transfectants were immunoprecipitated (IP) by anti-FLAG affinity agarose gel (upper panels), anti-Gα₁₆ or anti-Gα_z antiserum (middle panels). The immunoprecipitates were immunoblotted with anti-Gα₁₆, anti-Gα_z, or anti-FLAG antiserum. Aliquots of cell lysates were used to detect the expression levels of Gα₁₆, Gα_z and FLAG-TPR1 by western blot analysis (lower panels). Data shown represent one of three sets of immunoblots; two other sets yielded similar results.