An allosteric switch between the activation loop and a c-terminal palindromic phospho-motif controls c-Src function.
An allosteric switch between the activation loop and a c-terminal palindromic phospho-motif controls c-Src function.
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DOI：
10.1038/s41467-023-41890-7
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发表时间：
2023-10-17
影响因子：
16.6
通讯作者：
Plaza-Menacho, Ivan
中科院分区：
文献类型：
--
作者：
Cuesta-Hernandez, Hipolito Nicolas;Contreras, Julia;Soriano-Maldonado, Pablo;Sanchez-Wandelmer, Jana;Yeung, Wayland;Martin-Hurtado, Ana;Munoz, Ines G.;Kannan, Natarajan;Llimargas, Marta;Munoz, Javier;Plaza-Menacho, Ivan
自磷酸化控制蛋白激酶的离散功能和构象状态之间的过渡，但这一基本过程的结构和分子决定因素仍然不清楚。在这里，我们表明，C-末端Tyr 530是一个事实上的c-Src自磷酸化位点，具有缓慢的时间分辨率动力学和强大的分子间组分。相反，激活环Tyr 419经历更快的动力学和顺式至反式磷酸化开关，其控制c-末端Tyr 530自磷酸化、酶特异性和显著的c-Src非催化功能作为底物。与此一致，我们通过X射线晶体学可视化Tyr 530分子间自磷酸化的快照。在两个催化结构域的不对称排列中，底物分子上Tyr 530侧翼的C-末端回文磷酸基序与活性激酶的G-环接合，采用准备进入催化裂缝的位置。磷酸基序的扰动导致c-Src功能障碍，如病毒和结直肠癌（CRC）相关的c-末端缺失变体所示。我们表明，C-末端残基531至536所需的c-Src酪氨酸530自磷酸化，这种不利的影响是由底物分子抑制变构活性激酶。我们的工作揭示了激活和C-末端片段之间的串扰，控制底物和酶作用激酶在自磷酸化过程中的变构相互作用。蛋白激酶在不同构象状态之间的转换是由自磷酸化控制的。在这里，作者证明了c-末端Tyr 530是一个事实上的c-Src自磷酸化位点，并确定了一个关键的c-末端回文磷酸基序，它控制着自磷酸化过程中底物和酶作用激酶之间的相互作用。
Autophosphorylation controls the transition between discrete functional and conformational states in protein kinases, yet the structural and molecular determinants underlying this fundamental process remain unclear. Here we show that c-terminal Tyr 530 is a de facto c-Src autophosphorylation site with slow time-resolution kinetics and a strong intermolecular component. On the contrary, activation-loop Tyr 419 undergoes faster kinetics and a cis-to-trans phosphorylation switch that controls c-terminal Tyr 530 autophosphorylation, enzyme specificity, and strikingly, c-Src non-catalytic function as a substrate. In line with this, we visualize by X-ray crystallography a snapshot of Tyr 530 intermolecular autophosphorylation. In an asymmetric arrangement of both catalytic domains, a c-terminal palindromic phospho-motif flanking Tyr 530 on the substrate molecule engages the G-loop of the active kinase adopting a position ready for entry into the catalytic cleft. Perturbation of the phospho-motif accounts for c-Src dysfunction as indicated by viral and colorectal cancer (CRC)-associated c-terminal deleted variants. We show that c-terminal residues 531 to 536 are required for c-Src Tyr 530 autophosphorylation, and such a detrimental effect is caused by the substrate molecule inhibiting allosterically the active kinase. Our work reveals a crosstalk between the activation and c-terminal segments that control the allosteric interplay between substrate- and enzyme-acting kinases during autophosphorylation. Protein kinase transition between different conformational states is controlled by autophosphorylation. Here, the authors demonstrate that the c-terminal Tyr530 is a de facto c-Src autophosphorylation site and identify a critical c-terminal palindromic phospho-motif that controls the interplay between substrate and enzyme-acting kinases during autophosphorylation.
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影响因子：
3.3
作者：
Deminoff, Stephen J.;Ramachandran, Vidhya;Herman, Paul K.
通讯作者：
Herman, Paul K.
影响因子：
2.5
作者：
Cowieson, Nathan P.;Edwards-Gayle, Charlotte J. C.;Rambo, Robert P.
通讯作者：
Rambo, Robert P.
影响因子：
64.8
作者：
BAGRODIA, S;CHACKALAPARAMPIL, I;SHALLOWAY, D
通讯作者：
SHALLOWAY, D
影响因子：
64.8
作者：
Curtis, Christina;Shah, Sohrab P.;Chin, Suet-Feung;Turashvili, Gulisa;Rueda, Oscar M.;Dunning, Mark J.;Speed, Doug;Lynch, Andy G.;Samarajiwa, Shamith;Yuan, Yinyin;Graef, Stefan;Ha, Gavin;Haffari, Gholamreza;Bashashati, Ali;Russell, Roslin;McKinney, Steven;Langerod, Anita;Green, Andrew;Provenzano, Elena;Wishart, Gordon;Pinder, Sarah;Watson, Peter;Markowetz, Florian;Murphy, Leigh;Ellis, Ian;Purushotham, Arnie;Borresen-Dale, Anne-Lise;Brenton, James D.;Tavare, Simon;Caldas, Carlos;Aparicio, Samuel
通讯作者：
Aparicio, Samuel
影响因子：
64.5
作者：
IMAMOTO, A;SORIANO, P
通讯作者：
SORIANO, P