Insight into the C-terminal SH3 domain mediated binding of Drosophila Drk to Sos and Dos
Insight into the C-terminal SH3 domain mediated binding of Drosophila Drk to Sos and Dos
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DOI：
10.1016/j.bbrc.2022.08.007
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发表时间：
2022-08-08
影响因子：
3.1
通讯作者：
Ito, Yutaka
中科院分区：
文献类型：
--
作者：
Sayeesh, Pooppadi Maxin;Ikeya, Teppei;Ito, Yutaka
Drk是人类GRB2的果蝇同源物，通过其SH2结构域与seven - less （Sev）受体相互作用，而n端和c端SH3结构域（分别为Drk- nsh3和Drk- csh3）负责与seven - less之子（Sos）或seven - less之女（Dos）的富含脯氨酸基序（PRMs）相互作用。Drk-NSH3本身在折叠和未折叠状态之间具有构象平衡，并且通过与带有PxxPxR基序的富含脯氨酸的sos衍生肽的结合来稳定折叠状态。相反，Drk-CSH3应该结合Dos中的PxxxRxxKP基序。为了阐明两个SH3结构域之间的结构和功能差异，我们对Drk-CSH3进行了核磁共振研究。得到的溶液结构和n -15弛豫数据表明，Drk-CSH3由一个稳定的结构域组成。在RT环和疏水斑块周围通常发现大的化学位移扰动，而在Sos或dos衍生的肽中也有特征性的变化。Sos衍生的两个PxxPxR基序肽对Drk-CSH3具有较强的亲和力，表明Sos PRMs可以结合N-和C-SH3结构域。dos衍生的两个肽也可以结合Drk-CSH3，但亲和力要弱得多，这表明Dos-PRMs的任何合作结合都可能加强Drk-Dos的相互作用。核磁共振研究以及对接模拟为Drk中两个SH3结构域的生物学和生物物理功能提供了有价值的见解。(C) 2022作者。Elsevier Inc.出版。
Drk, a Drosophila homologue of human GRB2, interacts with Sevenless (Sev) receptor via its SH2 domain, while the N-and C-terminal SH3 domains (Drk-NSH3 and Drk-CSH3, respectively) are responsible for the interaction with proline-rich motifs (PRMs) of Son of sevenless (Sos) or Daughter of Sevenless (Dos). Drk-NSH3 on its own has a conformational equilibrium between folded and unfolded states, and the folded state is stabilised by the association with a Sos-derived proline-rich peptide with PxxPxR motif. In contrast, Drk-CSH3 is supposed to bind PxxxRxxKP motifs in Dos. Aiming at clarifying the structural and functional differences between the two SH3 domains, we performed NMR studies of Drk-CSH3. The resulting solution structure and the N-15-relaxation data showed that Drk-CSH3 consists of a stable domain. Large chemical shift perturbation was commonly found around the RT loop and the hydrophobic patch, while there were also changes that occur characteristically for Sos- or Dos-derived peptides. Sos-derived two peptides with PxxPxR motif showed stronger affinity to Drk-CSH3, indicating that the Sos PRMs can bind both N- and C-SH3 domains. Dos-derived two peptides could also bind Drk-CSH3, but with much weaker affinity, suggesting a possibility that any cooperative binding of Dos-PRMs may strengthen the Drk-Dos interaction. The NMR studies as well as the docking simulations provide valuable insights into the biological and biophysical functions of two SH3 domains in Drk. (C) 2022 The Authors. Published by Elsevier Inc.