Figure 1. Proteomic analysis of... | Rockefeller University Press

Figure 1.

$Proteomic analysis of cAMP-induced translocation. (A) Outline of the proteomic experiment for identification of polarity regulators. Red circles and blue triangles symbolize potential leading and lagging edge proteins, which transiently translocate to or dissociate from the plasma membrane upon stimulation, respectively, and hence accumulate differentially in the peripheral membrane protein fractions. (B) PHcrac-GFP translocation upon the addition of cAMP at time 0. Scale bar = 10 µm. (C) Cells expressing PHcrac-GFP were stimulated with cAMP, lysed at the indicated time points, and processed to obtain different membrane fractions, which were then probed with an anti-GFP antibody. PHcrac-GFP was found to be enriched in fractions collected at 10 and 20 s. CBB, Coomassie Brilliant Blue. (D and E) Plot of the translocation scores of 2,069 proteins identified in the MS analysis, with each dot representing a protein. Several known leading edge regulators are marked on the graph, and their respective translocation scores are listed in E. The translocation score of Leep1 is 0.69.$

Proteomic analysis of cAMP-induced translocation. (A) Outline of the proteomic experiment for identification of polarity regulators. Red circles and blue triangles symbolize potential leading and lagging edge proteins, which transiently translocate to or dissociate from the plasma membrane upon stimulation, respectively, and hence accumulate differentially in the peripheral membrane protein fractions. (B) PHcrac-GFP translocation upon the addition of cAMP at time 0. Scale bar = 10 µm. (C) Cells expressing PHcrac-GFP were stimulated with cAMP, lysed at the indicated time points, and processed to obtain different membrane fractions, which were then probed with an anti-GFP antibody. PHcrac-GFP was found to be enriched in fractions collected at 10 and 20 s. CBB, Coomassie Brilliant Blue. (D and E) Plot of the translocation scores of 2,069 proteins identified in the MS analysis, with each dot representing a protein. Several known leading edge regulators are marked on the graph, and their respective translocation scores are listed in E. The translocation score of Leep1 is 0.69.