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. Author manuscript; available in PMC: 2015 Dec 2.
Published in final edited form as: Structure. 2014 Dec 2;22(12):1844–1854. doi: 10.1016/j.str.2014.10.008

Figure 2.

Figure 2

Basal activity and DSF measurements of PLCβ3 variants. (A) Deletions within the X–Y linker, but not the Gαq binding site, alter basal activity. Increased activity is observed when the acidic stretch or entire linker is removed. Data represent at least five experiments performed in duplicate. (B) Addition of 5 mM IP3 to hyperactive PLCβ3 variants increases their thermal stability (see Supplemental Figure 3A). Tm values were determined by monitoring the increase in fluorescence of ANS as a function of temperature. See Supplemental Table 2. Data represent at least five experiments performed in triplicate. (C) Deletions including the acidic stretch show decreased thermal stability compared to variants containing intact X–Y linkers. The Tm of each PLCβ3 variant was determined with respect to PLCβ3-Δ847, used as a control on each plate (Supplemental Table 3). PLCβ3-Δ892 and its variants have higher thermal stability relative to PLCβ3-Δ847 variants due to Hα2′. Data represents at least four experiments performed in triplicate. In panels B and C, * = P≤0.5, ** = P≤0.01, *** = P≤0.001, **** =P≤0.0001.