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Peptide substrates for G protein‐coupled receptor kinase 2
Author(s) -
Asai Daisuke,
Toita Riki,
Murata Masaharu,
Katayama Yoshiki,
Nakashima Hideki,
Kang Jeong-Hun
Publication year - 2014
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2014.04.038
Subject(s) - g protein coupled receptor kinase , beta adrenergic receptor kinase , peptide , phosphorylation , kinase , biochemistry , receptor , amino acid , peptide sequence , chemistry , biology , g protein coupled receptor , gene
G protein‐coupled receptor kinases (GRKs) control the signaling and activation of G protein‐coupled receptors through phosphorylation. In this study, consensus substrate motifs for GRK2 were identified from the sequences of GRK2 protein substrates, and 17 candidate peptides were synthesized to identify peptide substrates with high affinity for GRK2. GRK2 appears to require an acidic amino acid at the −2, −3, or −4 positions and its consensus phosphorylation site motifs were identified as (D/E)X 1–3 ( S / T ), (D/E)X 1–3 ( S / T )(D/E), or (D/E)X 0–2 (D/E)( S / T ). Among the 17 peptide substrates examined, a 13‐amino‐acid peptide fragment of β‐tubulin (DEMEF T EAESNMN) showed the highest affinity for GRK2 ( K m , 33.9 μM; V max , 0.35 pmol min −1 mg −1 ), but very low affinity for GRK5. This peptide may be a useful tool for investigating cellular signaling pathways regulated by GRK2.