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Identification of arrestin elements mediating c‐Raf‐1 binding
Author(s) -
Coffa Sergio,
Song Xiufeng,
Gurevich Vsevolod V
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.710.1
Subject(s) - arrestin , microbiology and biotechnology , ask1 , g protein coupled receptor , mapk/erk pathway , kinase , signal transduction , protein kinase a , c raf , map kinase kinase kinase , chemistry , mitogen activated protein kinase kinase , biology
Arrestins regulate signaling and trafficking of G protein‐coupled receptor (GPCR). GPCR‐arrestin complexes initiate several signaling pathways, including the activation of mitogen activated protein kinases (MAPKs), such as extracellular signal regulated kinase 1/2 (ERK1/2) and c‐Jun N‐terminal kinase 3 (JNK3). Different MAPKs are involved in proliferation, survival, and apoptosis. Understanding of the structure and dynamics of arrestin‐MAPK complexes is necessary to design signaling‐biased arrestins to re‐direct MAPK signaling. We showed that c‐Raf1, MEK1, ERK2, ASK1, MKK4, and JNK3 interact with both domains of arrestin2 and arrestin3. Thus, three kinases in each cascade (c‐Raf‐1/MEK1/ERK2 or ASK1/MKK4/JNK3) are oriented in the same way parallel to the long axis of the arrestin molecule. We mutated residues on the non receptor‐binding surface conserved between arrestin2 and arrestin3. We found that the binding of arrestin2‐Arg307Ala mutant to c‐Raf‐1 is reduced by ~50%. Arg307Ala mutation does not affect the binding to phosphorylated receptor, MEK1, ERK2, and other MAPKs, ruling out gross folding defects. Thus, Arg307 plays a specific role in arrestin interaction with c‐Raf‐1. Arg307Ala mutant can disrupt the arrestin‐dependent ERK activation and will be a useful tool to study arrestin‐dependent MAPK activation. NIH grants GM077561 and GM081756 (VVG).