30 U87-MG MCF10A P-REX2a OHPH P-REX2a+ P-REX2a-N212A - PTEN PTEN DHPH (VS) Dutio

Question

30 U87-MG MCF10A P-REX2a OHPH P-REX2a+ P-REX2a-N212A - PTEN PTEN DHPH (VS) Dution Butler AKT 05 Molar Ratio (P-REX2a/PTEN) 1.5 MCF10A (reduced growth factors) 2) control 5-DHPH DM 3"A 1S.EB112?179 9 10M 6077 5.076 50.01-71.54 60 0.5 40 Day Fig. 2. Inhibition of PTEN phosphatase activity by P-REX2a. (A) Full-length P-REX2a, or a deletion of the DHPH domain (ADHPH), or the DHPH domain alone were added in the indicated molar volumes to 40 nM PTEN, purified from HEK293 cells, and phosphatase activity of PTEN was assayed with 20 ?M oo di-Ca-PIP3. This is a representative experiment error bars indicate standard deviation (n 3), "P

Explanation / Answer

A. Pten is a phosphatase and tension homologue, which exhibits phosphatase activity towards proteins and lipids. PREX2a is a PTEN inhibitor which is added in molar volumes to the solution of 40nm PTEN which is extracted from HEK293 cells. In the figure, phosphatase activity of PTEN is plotted against molar volume of PREX 2a to PTEN, where addition of PREX2a reduced PTEN phosphatase activity and error bar is also provided, elation buffer doesn’t show any reduction in activity.

B. Phosphatase activity of PTEN was plotted against volume of PREX2a added in the range of 0, 10 and 20 nm to estimate reduction of activity. Accordingly, increase in volume of PREX2a increase in reduction of phosphatase activity obviously.

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