Probing crystallin using serial UV/X-ray crystallography - Briony Yorke

Serial crystallography provides opportunities to investigate protein dynamics at high spatial and temporal resolution. We are applying this technique to the investigation of photostability and photodamage to proteins to identify conformational changes that occur in response to exposure to light. Ultraviolet radiation (UVR) is known to induce structural changes in proteins through two major pathways - primary photodamage caused by direct absorption of photons by aromatic amino acids and secondary photodamage through the formation of reactive oxygen species (ROS) which
contribute to protein oxidation. Both pathways are known to result in changes to the physical and chemical properties of proteins and enzymes, contributing to loss of stability, activity, and solubility. An unusual example of a UVR stable protein found in nature is Human Gamma-D crystallin (HGD), one of the major constituents of the human eye lens. Which has evolved to retain transparency over the course of a human life span in the absence of cellular regeneration and with daily exposure to UVR. Using serial crystallographic methods, we aim to understand the structural origins of HGD stability and the mechanisms of photodamage.