Real-time capture of domain movements during copper amine oxidase catalysis by mix-and-inject serial crystallography.

ABSTRACT: Protein dynamics play a crucial role in various physiological functions, including enzyme catalysis. To explore conformational changes during enzyme catalysis, we conduct mix-and-inject serial crystallography, an advanced technique to capture time-resolved protein structures in real time, using the microcrystals of bacterial copper amine oxidase containing a protein-derived quinone cofactor. Within 50 ms of mixing the microcrystals (<4 μm) with a preferred substrate (2-phenylethylamine) under anaerobic conditions (reductive half-reaction), we observe domain movements associated with substrate binding and formation of a metastable reaction intermediate, a product Schiff-base of the quinone cofactor. At 100-1000 ms after mixing, conformational transition from aminoresorcinol to the semiquino

SUBMITTER: Murakawa T 

PROVIDER: S-EPMC12714705 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature