Part One of the proposal is concerned with the fundamental structure of the TBM and aims to determine: (i) the impact of spacer molecule length on the dimensions of the bilayer and the water profile across the whole structure, & (ii) With a short spacer molecule is the membrane geometry altered? If so how, does the thickness of the bilayer change, the water profile or both ? Is the space above the short tether filled by solution or does the bilayer ¿collapse¿ into this space. Part Two will study the insertion of Chloride Intracellular Channel (CLIC1), specifically (i) the relative position and dimensions of the inserted CLIC1 within bilayer; and (ii) the amount of protein incorporated into the bilayer. Both questions will be approached using various ratios of D2O/H2O, and h- d-CLIC protein to define the protein location which will be compared with current spectroscppic insertion models.