Structural basis of dynamic P5CS filaments

The bifunctional enzyme triangle(1)- pyrroline- 5- carboxylate synthase (P5CS) is vital to the synthesis of proline and ornithine, playing an essential role in human health and agriculture. Pathogenic mutations in the P5CS gene (ALDH18A1) lead to neurocutaneous syndrome and skin relaxation connective tissue disease in humans, and P5CS deficiency seriously damages the ability to resist adversity in plants. We have recently found that P5CS forms cytoophidia in vivo and filaments in vitro. However, it is difficult to appreciate the function of P5CS filamentation without precise structures. Using cryo-electron microscopy, here we solve the structures of Drosophila full-length P5CS in three states at resolution from 3.1 to 4.3 A. We observe distinct ligand-binding states and conformational changes for the GK and GPR domains, respectively. Divergent helical filaments are assembled by P5CS tetramers and stabilized by multiple interfaces. Point mutations disturbing those interfaces prevent P5CS filamentation and greatly reduce the enzymatic activity. Our findings reveal that filamentation is crucial for the coordination between the GK and GPR domains, providing a structural basis for the catalytic function of P5CS filaments.

Automated summary

The bifunctional enzyme P5CS plays a vital role in proline and ornithine synthesis and its mutations can lead to human diseases and plant resistance impairment. We have discovered that P5CS forms cytoophidia in vivo and filaments in vitro, and have solved the structures of Drosophila full-length P5CS using cryo-electron microscopy. The filamentation of P5CS is crucial for the coordination between different domains and provides a structural basis for its catalytic function.