Publication
A multi-reservoir extruder for time-resolved serial protein crystallography and compound screening at X-ray free-electron lasers
Publisher:
Research Square Platform LLC
Date:
09-03-2023
DOI:
10.21203/RS.3.RS-2621995/V1
Abstract: Serial crystallography at X-ray free-electron lasers (XFELs) permits the determination of radiation-damage free static as well as time-resolved protein structures at room temperature. Efficient s le delivery is a key factor for such experiments. Here, we describe a multi-reservoir, high viscosity extruder as a step towards automation of s le delivery at XFELs. Compared to a standard single extruder, s le exchange time was halved and the workload of users was greatly reduced. In-built temperature control of s les facilitated optimal extrusion and offered a path forward for the observation of temperature-dependent structural changes. After commissioning the device with lysozyme crystals, we collected time-resolved data using crystals of a membrane-bound, light-driven sodium pump. Static data were also collected from the soluble protein tubulin that was soaked with a series of small molecule drugs. Using these data, we identified low occupancy (as little as 30%) ligands using a minimal amount of data from a serial crystallography experiment, a result that could be exploited for structure-based drug design.