A method to stabilize reduced and/or gas-treated protein crystals by flash-cooling under a controlled atmosphere

A customized glove box for protein crystallization under a controlled atmosphere is described along with a cryogenic technique adapted to freeze protein crystals inside the glove box and a very simple device for studying gas-protein complexes in the crystalline state at cryogenic temperatures. Using these techniques different redox states of oxygen-sensitive crystalline proteins have been stabilized and the interaction of hydrogenase with Xe, a model for the much lighter substrate molecular hydrogen, has been studied.