Aconitase Regulation of Erythropoiesis Correlates with a Novel Licensing Function in Erythropoietin-Induced ERK Signaling
Figure 7
Identification of an ERK-aconitase protein complex that is disrupted by aconitase inhibition.
(A) Assessment of ERK-aconitase interactions by coexpression followed by coimmunoprecipitation. ERK2 and FLAG-tagged cytosolic aconitase (cAcon-FLAG) or mitochondrial aconitase (mAcon-FLAG) were coexpressed in HEK293T cells ±500 µM FA. Immunoprecipitation (IP) with FLAG beads was followed by immunoblotting (IB) for total ERK and for the FLAG epitope. (B) Role of ERK D-domain interacting motif in ERK2 binding to mitochondrial aconitase. mAcon-FLAG was coexpressed with ERK2 WT or D319N in HEK293T cells ±500 µM FA, ±100 ng/ml EGF, followed by IP and IB as in panel A.