Howard James A, Hok Lucija, Cate Richard L, Sanford Nathaniel J, Hart Kaitlin N, Leach Edmund A E, Bruening Alena S, Nagykery Nicholas, Donahoe Patricia K, Pépin David, Thompson Thomas B
Department of Pharmacology, Physiology, and Neurobiology, University of Cincinnati, Cincinnati, OH 45267.
Department of Molecular & Cellular Biosciences, University of Cincinnati, Cincinnati, OH 45267.
Proc Natl Acad Sci U S A. 2025 Jan 21;122(3):e2418088122. doi: 10.1073/pnas.2418088122. Epub 2025 Jan 13.
TGFβ family ligands are synthesized as precursors consisting of an N-terminal prodomain and C-terminal growth factor (GF) signaling domain. After proteolytic processing, the prodomain typically remains noncovalently associated with the GF, sometimes forming a high-affinity latent procomplex that requires activation. For the TGFβ family ligand anti-Müllerian hormone (AMH), the prodomain maintains a high-affinity interaction with its GF that does not render it latent. While the prodomain can be displaced by the type II receptor, AMHR2, the nature of the GF:prodomain interaction and the mechanism of prodomain displacement by AMHR2 are currently unknown. We show here that the AMH prodomain exhibits an atypical two-domain structure, containing a dimerizing and a GF-binding domain connected through a flexible linker. Cryo-EM and genomic analyses show that the distinctive GF-binding domain, the result of an exon insertion 450 Mya, comprises a helical bundle and a belt-like structure which interact with the GF at the type II and I receptor binding sites, respectively. The dimerizing domain, which adopts a TGFβ-like propeptide fold, covalently connects two prodomains through intermolecular disulfide bonds. Disease mutations map to both the GF-binding and dimerization domains. Our results support a model where AMHR2 displaces the helical bundle and induces a conformational change in the GF, followed by release of the prodomain and engagement of the type I receptor. Collectively, this study shows that the AMH prodomain has evolved an atypical binding interaction with the GF that favors, without disrupting signaling, the maintenance of a noncovalent complex until receptors are engaged.
转化生长因子β(TGFβ)家族配体作为前体被合成,该前体由N端前结构域和C端生长因子(GF)信号结构域组成。经过蛋白水解加工后,前结构域通常仍与GF非共价结合,有时会形成需要激活的高亲和力潜伏前复合物。对于TGFβ家族配体抗苗勒管激素(AMH),前结构域与其GF保持高亲和力相互作用,不会使其处于潜伏状态。虽然前结构域可以被II型受体AMHR2取代,但GF与前结构域相互作用的性质以及AMHR2取代前结构域的机制目前尚不清楚。我们在此表明,AMH前结构域呈现出一种非典型的双结构域结构,包含通过柔性接头连接的二聚化结构域和GF结合结构域。冷冻电镜和基因组分析表明,独特的GF结合结构域是4.5亿年前外显子插入的结果,它由一个螺旋束和一个带状结构组成,分别在II型和I型受体结合位点与GF相互作用。二聚化结构域采用TGFβ样前肽折叠,通过分子间二硫键共价连接两个前结构域。疾病突变映射到GF结合结构域和二聚化结构域。我们的结果支持这样一个模型,即AMHR2取代螺旋束并诱导GF发生构象变化,随后前结构域释放,I型受体结合。总的来说,这项研究表明,AMH前结构域与GF进化出了一种非典型的结合相互作用,这种相互作用有利于在不破坏信号传导的情况下维持非共价复合物,直到受体结合。