美国诺华生物医学研究（NBR）David J. Rowlands研究团队开发出变构性HECT E3连接酶抑制的治疗潜力。这一研究成果发表在2025年4月2日出版的国际学术期刊《细胞》上。
在一个大型的、无偏倚的生化筛选中，研究小组发现抑制剂结合了一个远离E6相关蛋白C末端结构域（HECT） E3连接酶的催化半胱氨酸的隐腔，SMAD泛素调节因子1（SMURF1）。结构和生化分析以及工程逃逸突变表明，这些抑制剂通过在保守的甘氨酸铰链上延伸α螺旋来限制基本的催化运动。SMURF1水平在肺动脉高压（PAH）中升高，PAH是一种由骨形态发生蛋白受体-2（BMPR2）突变引起的疾病。该团队证明SMURF1抑制可阻止BMPR2泛素化，使骨形态发生蛋白（BMP）信号正常化，恢复肺血管细胞稳态，并逆转已建立的实验性多环芳烃的病理。利用这种深入的机制理解，小组进行了基于计算机机器学习的筛选，以确定原型HECT E6AP的抑制剂，并在体外确认甘氨酸铰链依赖性变构活性。抑制HECTs和其他甘氨酸铰链蛋白开辟了一个新的药物空间。
据介绍，靶向泛素E3连接酶在治疗上具有吸引力；然而，活性位点口袋的缺失阻碍了识别抑制剂的计算方法。
附：英文原文
Title: Therapeutic potential of allosteric HECT E3 ligase inhibition
Author: Alexander M.K. Rothman, Amir Florentin, Florence Zink, Catherine Quigley, Olivier Bonneau, Rene Hemmig, Amanda Hachey, Tomas Rejtar, Maulik Thaker, Rishi Jain, Shih-Min Huang, Daniel Sutton, Jan Roger, Ji-Hu Zhang, Sven Weiler, Simona Cotesta, Johannes Ottl, Salil Srivastava, Alina Kordonsky, Reut Avishid, Elon Yariv, Ritu Rathi, Oshrit Khvalevsky, Thomas Troxler, Sarah K. Binmahfooz, Oded Kleifeld, Nicholas W. Morrell, Marc Humbert, Matthew J. Thomas, Gabor Jarai, Rohan E.J. Beckwith, Jennifer S. Cobb, Nichola Smith, Nils Ostermann, John Tallarico, Duncan Shaw, Sabine Guth-Gundel, Gali Prag, David J. Rowlands
Issue&Volume: 2025-04-02
Abstract: Targeting ubiquitin E3 ligases is therapeutically attractive; however, the absence of an active-site pocket impedes computational approaches for identifying inhibitors. In a large, unbiased biochemical screen, we discover inhibitors that bind a cryptic cavity distant from the catalytic cysteine of the homologous to E6-associated protein C terminus domain (HECT) E3 ligase, SMAD ubiquitin regulatory factor 1 (SMURF1). Structural and biochemical analyses and engineered escape mutants revealed that these inhibitors restrict an essential catalytic motion by extending an α helix over a conserved glycine hinge. SMURF1 levels are increased in pulmonary arterial hypertension (PAH), a disease caused by mutation of bone morphogenetic protein receptor-2 (BMPR2). We demonstrated that SMURF1 inhibition prevented BMPR2 ubiquitylation, normalized bone morphogenetic protein (BMP) signaling, restored pulmonary vascular cell homeostasis, and reversed pathology in established experimental PAH. Leveraging this deep mechanistic understanding, we undertook an in silico machine-learning-based screen to identify inhibitors of the prototypic HECT E6AP and confirmed glycine-hinge-dependent allosteric activity in vitro. Inhibiting HECTs and other glycine-hinge proteins opens a new druggable space.
DOI: 10.1016/j.cell.2025.03.001
Source: https://www.cell.com/cell/abstract/S0092-8674(25)00274-0