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. 2019 Jun 3;116(25):12500–12505. doi: 10.1073/pnas.1820131116

Fig. 2.

Fig. 2.

UBE3A ubiquitinates PTPA. (A) GST pull-down assay to validate the direct interaction between UBE3A and PTPA proteins in vitro. (B) PTPA was ubiquitinated by UBE3A but not its ligase-dead mutant Ube3a-C838A in HEK293T cells. Immunoblots were incubated with anti-HA antibodies. *, nonspecific bands. (C) In vitro ubiquitination of PTPA mediated by UBE3A analyzed by immunoblotting. (D) In vitro ubiquitination of PTPA using WT and Ube3am−/p+ mice brain lysates. (E) The lysine residues in PTPA ubiquitinated by UBE3A analyzed by LC-MS/MS. Rectangles in green indicate ATP binding sites. (F) PTPA mutation (4KR) blocked UBE3A-mediated PTPA degradation. Three independent experiments were performed.