Mix-and-extrude using 3D printed nozzles for time-resolved membrane protein crystallography

Abstract

AbstractTime-resolved crystallography enabled the visualization of protein molecular motion during reaction. Currently, light is the most prevalent reaction initiator, but the number of light-activated proteins is extremely limited. Instead, more biological reactions are triggered by interaction with ligands. To examine the molecular action of these mixing-initiated reactions by X-ray diffraction, a sample delivery method that allows for diffusive mixing of crystals with ligands in a short amount of time corresponding to the reaction time to be studied is needed. This paper describes the mix-and-extrude nozzle for delivering membrane proteins crystallized in lipid cubic phase and ligands, along with its preliminary characterization and outlook for future developments.Synopsis3D printed mixing-HVE devices address time-resolved membrane protein crystallography challenges via compact dual-flow LCP injection.