Raman spectra measurements on DEPC liposome and cell membrane of living neuron under xenon pressure

The Raman spectra of liposomes were measured under xenon pressures and low temperatures to observe the spectra changes accompanying the gel to liquid crystalline phase transition of the liposomes. C–H stretching bonds of the lipids in the liposome were slightly red shifted at approximately 285 K and atmospheric pressure, which coincided well with the phase transition condition. This Raman peak shift was observed at lower temperatures and related linearly to the xenon pressures. The xenon pressure dependence on the phase transition temperature was in good agreement with the DSC measurements, and the red shifts of Raman peaks supported the molecular mechanism of interaction between xenon and phospholipid bilayers suggested by the MD simulations. The phase transition measurements under xenon pressure with the microscopic Raman spectroscopy were applied to cultured neuronal networks to observe the interaction of dissolved xenon with the cell membrane and the surrounding water.

Freeze-Trapping Isomorphous Xenon Derivatives of Protein Crystals

A simple and efficient method to prepare isomorphous derivatives of protein crystals with xenon as a heavy atom is described. The method consists of exposing a crystal to xenon gas of pressures above 5 atm (~ 0.5 MPa) for several minutes and subsequently shock-freezing the crystal to immobilize the xenon dissolved in the mother liquor and bound to the protein. Diffraction data can the be collected with the established techniques of protein cryocrystallography. Two types of high-pressure device are described to expose a protein crystal to the required xenon pressure, permitting rapid freeze-quenching after xenon exposure. One of these devices can be used for gas pressures up to and exceeding 5.0 MPa, with a gas consumption of a few millilitres of uncompressed gas. The technique has been tested with monoclinic crystals of sperm-whale metmyoglobin, which has four xenon binding sites. The results of these experiments are described and discussed. Potential applications of this technique include-besides the classical multiwavelength anomalous diffraction (MAD) or single isomorphous replacement with anomalous scattering (SIRAS) experiments-the derivation of low-angle phase information by modifying the electron density of the solvent regions within the crystal.