Figure 2
From: Neuralized family member NEURL1 is a ubiquitin ligase for the cGMP-specific phosphodiesterase 9A
NEURL1-PDE9A interaction is mediated by the NHR1 and NHR2 domains of NEURL1 and the catalytic and regulatory domains of PDE9A. (A) Schematic diagram of the various NEURL1 and PDE9A constructs used. (B) NEURL1-Flag, NEURL1-Rm-Flag Flag (encoding the ubiquitin ligase defective protein), Flag-NHR2 and PDE9A-E2 were expressed in various combinations in HEK293-FT cells. (C) PDE9A-V5His protein was expressed alone or in combination with either EGFP-NHR1 + NHR2 or NHR1-EGFP in HEK293-FT cells. (D) PDE9A-cat-E2 or PDE9A-reg-E2 (encoding the catalytic and regulatory domains of PDE9A, respectively) were expressed alone or together with Flag-NEURL1 in HEK293-FT cells. In B-D the proteins were immunoprecipitated and immunoblotted using the indicated antibodies. NHR, Neuralized Homology Repeat; Rm, mutant RING domain; cat, catalytic domain; reg, regulatory domain; IB, immunoblot; IP, immunoprecipitation.
