Kinase and C-domains are the key arrestin-binding elements in ASK1.
A, Constructs containing different domains of ASK1: N domain
(ASK1-N), N and Kinase domains (ASK1-(N+K)), C domain (ASK1-C), C and kinase
domains (ASK1-(C+K)). B and C, COS-7 cells were transfected
with HA-tagged ASK1 constructs shown in panel A, together with
FLAG-arrestin3 (FLAG-Arr3) (panel B), or HA-ASK1-(C+K) with
full-length arrestin3 (Arr3) or its N- (Arr3-N) and C-domain (Arr3-C)
(panel C), respectively. Immunoprecipitation and Western blot
analysis were performed as described in the legend to
Fig. 2A.