Structure guided studies of the interaction between PTP1B and JAK.
Structure guided studies of the interaction between PTP1B and JAK.
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DOI：
10.1038/s42003-023-05020-9
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发表时间：
2023-06-14
影响因子：
5.9
通讯作者：
Babon, Jeffrey J.
中科院分区：
文献类型：
--
作者：
Morris, Rhiannon;Keating, Narelle;Tan, Cyrus;Chen, Hao;Laktyushin, Artem;Saiyed, Tamanna;Liau, Nicholas P. D.;Nicola, Nicos A.;Tiganis, Tony;Kershaw, Nadia J.;Babon, Jeffrey J.
蛋白酪氨酸磷酸酶1B(PTP1B)是一种典型的蛋白酪氨酸磷酸酶，在多种由激酶驱动的信号通路中起着重要的调节作用。PTP1B显示出对双磷酸化底物的偏好。在这里，我们确定PTP1B是一种IL-6的抑制剂，并表明在体外，它可以使JAK家族的所有四个成员去磷酸化。为了详细了解JAK去磷酸化的分子机制，我们对去磷酸化反应进行了结构和生化分析。我们鉴定了一个捕获产品的PTP1B突变体，它允许可视化反应的酪氨酸和磷酸产物，以及一个底物捕获突变体，与之前描述的相比，失败率大大降低。后一个突变体被用来确定与酶活性部位结合的双磷酸化JAK多肽的结构。这些结构表明，与IRK的类似区域相反，下游的磷酸酪氨酸优先与活性部位结合。生化分析证实了这一偏好。在这种结合模式下，先前确定的第二个芳基结合部位保持未被占据，并且非底物磷酸酪氨酸与Arg47结合。这种精氨酸的突变打乱了对下游磷酸酪氨酸的偏好。这项研究揭示了PTP1B如何与不同底物相互作用的可塑性。磷酸酶和免疫检查点PTP1B的结构和生化特征揭示了PTBP1B/JAK相互作用的分子基础和PTP1B识别底物的可塑性。
Protein Tyrosine Phosphatase 1B (PTP1B) is the prototypical protein tyrosine phosphatase and plays an essential role in the regulation of several kinase-driven signalling pathways. PTP1B displays a preference for bisphosphorylated substrates. Here we identify PTP1B as an inhibitor of IL-6 and show that, in vitro, it can dephosphorylate all four members of the JAK family. In order to gain a detailed understanding of the molecular mechanism of JAK dephosphorylation, we undertook a structural and biochemical analysis of the dephosphorylation reaction. We identified a product-trapping PTP1B mutant that allowed visualisation of the tyrosine and phosphate products of the reaction and a substrate-trapping mutant with a vastly decreased off-rate compared to those previously described. The latter mutant was used to determine the structure of bisphosphorylated JAK peptides bound to the enzyme active site. These structures revealed that the downstream phosphotyrosine preferentially engaged the active site, in contrast to the analogous region of IRK. Biochemical analysis confirmed this preference. In this binding mode, the previously identified second aryl binding site remains unoccupied and the non-substrate phosphotyrosine engages Arg47. Mutation of this arginine disrupts the preference for the downstream phosphotyrosine. This study reveals a previously unappreciated plasticity in how PTP1B interacts with different substrates. Structural and biochemical characterization of the phosphastase and immune checkpoint PTP1B reveals the molecular basis of PTBP1B/JAK interaction and plasticity of substrate recognition by PTP1B.
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影响因子：
37.8
作者：
Lanahan AA;Lech D;Dubrac A;Zhang J;Zhuang ZW;Eichmann A;Simons M
通讯作者：
Simons M
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
11.8
作者：
Cheng, A;Uetani, N;Tremblay, ML
通讯作者：
Tremblay, ML
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
DOI：
10.1155/2011/376543
发表时间：
2011
期刊：
Journal of signal transduction
影响因子：
--
作者：
Aoudjit L;Jiang R;Lee TH;New LA;Jones N;Takano T
通讯作者：
Takano T