Peptide substrates for G protein-coupled receptor kinase 2
Peptide substrates for G protein-coupled receptor kinase 2
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DOI：
10.1016/j.febslet.2014.04.038
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发表时间：
2014-06-13
期刊：
影响因子：
3.5
通讯作者：
Kang, Jeong-Hun
中科院分区：
文献类型：
--
作者：
Asai, Daisuke;Toita, Riki;Kang, Jeong-Hun
G蛋白偶联受体激酶（GRIC）通过磷酸化控制G蛋白偶联受体的信号传导和活化。在这项研究中，GRK 2的共识底物基序从GRK 2蛋白底物的序列中确定，并合成了17个候选肽，以确定肽底物与GRK 2的高亲和力。GRK 2似乎需要在-2、-3或-4位置处的酸性氨基酸，并且其共有磷酸化位点基序被鉴定为（D/ E）X1-3（S/T）、（D/E）X1-3（S/T）（D/E）或（D/E）X 0 -2（D/E）（S/T）。在检测的17种肽底物中，β-微管蛋白的13个氨基酸的肽片段（DEMEFTEAESNMN）显示出对GRK 2的最高亲和力（K-m，33.9 μ M; V-max，0.35 pmol min（-1）mg（-1）），但对GRIC 5的亲和力非常低。该肽可能是研究GRK 2调控的细胞信号通路的有用工具。(C)2014年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
G protein-coupled receptor kinases (GRICs) 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 beta-tubulin (DEMEFTEAESNMN) showed the highest affinity for GRK2 (K-m, 33.9 mu M; V-max, 0.35 pmol min(-1) mg(-1)), but very low affinity for GRIC5. This peptide may be a useful tool for investigating cellular signaling pathways regulated by GRK2. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.