TY - JOUR
T1 - Peptide substrates for G protein-coupled receptor kinase 2
AU - Asai, Daisuke
AU - Toita, Riki
AU - Murata, Masaharu
AU - Katayama, Yoshiki
AU - Nakashima, Hideki
AU - Kang, Jeong Hun
N1 - Funding Information:
The authors thank Ms. Sigemi Terakubo and Ms. Ninyo Okamura for technical assistance. This work was financially supported by a Grant-in-aid for Scientific Research (A) (KAKENHI Grant No. 24245015 ) and (B) (KAKENHI Grant No. 23310085 ), and a Grant-in-aid for Young Scientist (B) (KAKENHI Grant No. 24790052 ) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan.
PY - 2014/6/13
Y1 - 2014/6/13
N2 - 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)X1-3(S/T), (D/E)X1-3(S/T)(D/E), or (D/E)X0-2(D/E)(S/T). Among the 17 peptide substrates examined, a 13-amino-acid peptide fragment of β-tubulin (DEMEFTEAESNMN) showed the highest affinity for GRK2 (Km, 33.9 μM; Vmax, 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.
AB - 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)X1-3(S/T), (D/E)X1-3(S/T)(D/E), or (D/E)X0-2(D/E)(S/T). Among the 17 peptide substrates examined, a 13-amino-acid peptide fragment of β-tubulin (DEMEFTEAESNMN) showed the highest affinity for GRK2 (Km, 33.9 μM; Vmax, 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.
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U2 - 10.1016/j.febslet.2014.04.038
DO - 10.1016/j.febslet.2014.04.038
M3 - Article
C2 - 24813628
AN - SCOPUS:84902173987
SN - 0014-5793
VL - 588
SP - 2129
EP - 2132
JO - FEBS Letters
JF - FEBS Letters
IS - 13
ER -