Grant: Research Tools and Instruments - Category 1 (<$150,000)

We are requesting funds to purchase an ultracentrifuge and rotors for cellular subfractionation. The new ultracentrifuge will replace a much older (~25 years) model that has been the work horse in our Department. Empirical reductionism has for many years been a central and productive approach to cell biology and biochemistry. In this method, complex systems are broken down into their components with the hope that by understanding the components some insight will be gained into the more complex system. One classic way of separating cells into their components remains the spinning of samples at high speed. Special instruments, called ultracentrifuges, are required to do this at very high speeds (generating forces in excess of 500 000 times that of gravity). While cells are the basic units of life they are themselves quite complicated and are defined by multiple subcellular compartments. Each of these compartments carries out specific and defined processes. In order to study the proteins and processes taking place in these compartments it is necessary to separate them from the other components of the cell. They can be separated from one another based on their densities by centrifugation. This proposal is for an ultracentrifuge and the necessary rotors to carry out these types of cellular subfractionation experiments. While the experiments described here involve a number of different organisms (ranging from bacteria to fungi to plants) and subcellular fractions of interest, they are all related by the common thread that they utilize or require ultracentrifugation to be carried out. The progress of the research described here hinges on access to an ultracentrifuge to carry out this cellular subfractionation.