Observation of substrate diffusion and ligand binding in enzyme crystals using high-repetition-rate mix-and-inject serial crystallography

Medientyp: E-Artikel

Titel: Observation of substrate diffusion and ligand binding in enzyme crystals using high-repetition-rate mix-and-inject serial crystallography

Beteiligte: Pandey, Suraj; Calvey, George; Katz, Andrea M.; Malla, Tek Narsingh; Koua, Faisal H. M.; Martin-Garcia, Jose M.; Poudyal, Ishwor; Yang, Jay-How; Vakili, Mohammad; Yefanov, Oleksandr; Zielinski, Kara A.; Bajt, Sasa; Awel, Salah; Doerner, Katarina; Frank, Matthias; Gelisio, Luca; Jernigan, Rebecca; Kirkwood, Henry; Kloos, Marco; Koliyadu, Jayanath; Mariani, Valerio; Miller, Mitchell D.; Mills, Grant; Nelson, Garrett; [...]

Erschienen: International Union of Crystallography (IUCr), 2021

Sprache: Nicht zu entscheiden

DOI: 10.1107/s2052252521008125

ISSN: 2052-2525

Entstehung:

Anmerkungen:

Beschreibung: Here, we illustrate what happens inside the catalytic cleft of an enzyme when substrate or ligand binds on single-millisecond timescales. The initial phase of the enzymatic cycle is observed with near-atomic resolution using the most advanced X-ray source currently available: the European XFEL (EuXFEL). The high repetition rate of the EuXFEL combined with our mix-and-inject technology enables the initial phase of ceftriaxone binding to the Mycobacterium tuberculosis β-lactamase to be followed using time-resolved crystallography in real time. It is shown how a diffusion coefficient in enzyme crystals can be derived directly from the X-ray data, enabling the determination of ligand and enzyme–ligand concentrations at any position in the crystal volume as a function of time. In addition, the structure of the irreversible inhibitor sulbactam bound to the enzyme at a 66 ms time delay after mixing is described. This demonstrates that the EuXFEL can be used as an important tool for biomedically relevant research.

Zugangsstatus: Freier Zugang

Nur in Feld suchen:

Zuletzt gesuchte Begriffe: