Figure 4.
HDX-MS analysis of p110γ-p84 activation by membrane-localized HRas and Gβγ and comparison with p110γ-p101
(A–C) Significant HDX-MS differences in the p110γ and p84 subunits between (A) plasma membrane mimic vesicles and plasma membrane mimic vesicles with 3 μM GTPγS-loaded lipidated HRas, (B) plasma membrane mimic vesicles and plasma membrane mimic vesicles with 3 μM Gβγ, and (C) plasma membrane mimic vesicles with 3 μM GTPγS-loaded lipidated HRas and plasma membrane mimic vesicles with both HRas and Gβγ (3 μM) are mapped on the structure of p110γ-p84 according to the legend in (A). A cartoon model is shown to the right with differences annotated. The sum of the number of deuteron difference is shown for p110γ, with red dots representing peptides showing statistically significant differences. Error is shown as the sum of the standard deviations across all time points (n = 3 for each time point).
(D) Significant HDX-MS differences in the p110γ and p101 subunits between plasma membrane mimic vesicles and plasma membrane mimic vesicles with Gβγ mapped on the structure of p110γ-p101 (PDB: 7MEZ) according to the legend. A cartoon model is shown to the right with differences annotated.
The HDX-MS data in (D) are from our previous study30 and are provided as a comparison with (B).