Probing the modulation of enzyme kinetics by multi-temperature, time-resolved serial crystallography

We present an environmental enclosure for fixed-target serial crystallography, enabling X-ray diffraction experiments in a temperature window from below 10 °C to above 70 °C - a universal parameter of protein function. Via 5D-SSX time-resolved experiments can now be carried out at physiological temperatures, providing fundamentally new insights into protein function. We show temperature-dependent modulation of turnover kinetics for the mesophilicβ-lactamase CTX-M-14 and for the hyperthermophilic enzyme xylose isomerase.