以及谷胱甘肽 S-转移酶与丝裂原活化蛋白激酶 8 和凋亡信号调节激酶 1 的蛋白-蛋白相互作用的研究。

and investigations on the protein-protein interactions of glutathione S-transferases with mitogen-activated protein kinase 8 and apoptosis signal-regulating kinase 1.

机构信息

Onco-Haematology Unit, Department of Paediatrics, Obstetrics and Gynaecology, Geneva University Hospitals, Geneva, Switzerland.

Research Platform on Pediatric Onco-Hematology, Department of Paediatrics, Obstetrics and Gynaecology, Faculty of Medicine, University of Geneva, Geneva, Switzerland.

出版信息

J Biomol Struct Dyn. 2022 Feb;40(3):1430-1440. doi: 10.1080/07391102.2020.1827036. Epub 2020 Sep 30.

DOI:10.1080/07391102.2020.1827036

PMID:32996404

Abstract

Cytosolic glutathione -transferase (GST) enzymes participate in several cellular processes in addition to facilitating glutathione conjugation reactions that eliminate endogenous and exogenous toxic compounds, especially electrophiles. GSTs are thought to interact with various kinases, resulting in the modulation of apoptotic processes and cellular proliferation. The present research used a combination of and studies to investigate protein-protein interactions between the seven most abundant cytosolic GSTs-GST alpha-1 (GST-A1), GST alpha-2 (GST-A2), GST mu-1 (GST-M1), GST mu-2 (GST-M2), GST mu-5 (GST-M5), GST theta-1 (GST-T1) and GST pi-1 (GST-P1)-and Mitogen-activated protein kinase 8 (MAPK8) and Apoptosis signal-regulating kinase 1 (ASK1). MAPK8 and ASK1 were chosen as this study's protein interaction partners because of their predominant role in electrophile or cytokine-induced stress-mediated apoptosis, inflammation and fibrosis. The highest degree of sequence homology or sequence similarity was observed in two GST subgroups: the GST-A1, GST-A2 and GST-P1 isoforms constituted subgroup1; the GST-M1, GST-M2 and GST-M5 isoforms constituted subgroup 2. The GST-T1 isoform diverged from these isoforms. investigations revealed that GST-M1 showed a significantly higher binding affinity to MAPK8, and its complex was more structurally stable than the other isoforms, in the order GST-M1 > GST-M5 > GST-P1 > GST-A2 > GST-A1 > GST-M2 > GST-T1. Similarly, GST-A1, GST-P1 and GST-T1 actively interacted with ASK1, and their structural stability was also better, in the order GST-T1 > GST-A1 > GST-P1 > GST-A2 > GST-M5 > GST-M1 > GST-M2. To validate results, we performed crosslinking and mass spectroscopy experiments. Results indicated that GST-M1 interacted with GST-T1 to form heterodimers and confirmed the predicted interaction between GST-M1 and MAPK8.Communicated by Ramaswamy H. Sarma.

摘要

细胞溶质谷胱甘肽-S-转移酶（GST）酶除了促进谷胱甘肽缀合反应以消除内源性和外源性有毒化合物（特别是亲电体）外，还参与许多细胞过程。GST 被认为与各种激酶相互作用，从而调节细胞凋亡过程和细胞增殖。本研究使用和研究相结合的方法，研究了七种最丰富的细胞溶质 GST-GSTα-1（GST-A1）、GSTα-2（GST-A2）、GSTMμ-1（GST-M1）、GSTMμ-2（GST-M2）、GSTMμ-5（GST-M5）、GSTθ-1（GST-T1）和 GSTπ-1（GST-P1）-与丝裂原活化蛋白激酶 8（MAPK8）和凋亡信号调节激酶 1（ASK1）之间的蛋白质-蛋白质相互作用。选择 MAPK8 和 ASK1 作为本研究的蛋白质相互作用伙伴，是因为它们在亲电体或细胞因子诱导的应激介导的细胞凋亡、炎症和纤维化中起着主要作用。在两个 GST 亚组中观察到最高程度的序列同源性或序列相似性：GST-A1、GST-A2 和 GST-P1 同工型构成亚组 1；GSTM1、GSTM2 和 GSTM5 同工型构成亚组 2。GST-T1 同工型与这些同工型不同。研究表明，GST-M1 与 MAPK8 的结合亲和力明显更高，其复合物的结构稳定性也优于其他同工型，顺序为 GST-M1>GST-M5>GST-P1>GST-A2>GST-A1>GST-M2>GST-T1。同样，GST-A1、GST-P1 和 GST-T1 与 ASK1 积极相互作用，并且它们的结构稳定性也更好，顺序为 GST-T1>GST-A1>GST-P1>GST-A2>GST-M5>GST-M1>GST-M2。为了验证结果，我们进行了交联和质谱实验。结果表明，GST-M1 与 GST-T1 相互作用形成异二聚体，并证实了 GST-M1 与 MAPK8 之间的预测相互作用。由 Ramaswamy H. Sarma 传达。