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

All living things have specialized cells, and within these cells, there are macromolecules (proteins, enzymes, nucleic acids) that carry out specific biological functions. The research groups of Drs Wolthers, Little and Klegeris require an ultracentrifuge for isolating these macromolecules. Isolation of the macromolecules allows determination of their composition, structure and biological functions. Dr. Wolthers will use the ultracentrifuge to isolate two types of enzymes, cytochrome P450 monooxygenase and its reducing partner, cytochrome P450 reductase. These enzymes are tethered to the membrane, or the outer surface of the cell. These two enzymes perform difficult reactions with exquisite control and are increasingly used by industry to manufacture pharmaceuticals and other complex chemicals. Her lab will study these enzymes to optimize their capacity to produce a range of complex chemicals. An ultracentrifuge is required to isolate these membrane proteins as it is capable of applying sufficient centrifugal force to separate the membranes from the soluble components of the cell. Dr. Little will use the ultracentrifuge to isolate a subclass of circulating microvesicles called exosomes. Exosomes are released from parent cells and circulate in the blood carrying distinct proteins and nucleic acids. Isolation and purification of microvesicles will allow his laboratory to characterize their cargo to reveal how they may play a novel role in inter-organ communication during physiological stress such as exercise. Dr. Klegeris' laboratory will use the ultracentrifuge to separate various components of cells in order to study proteins as they move from one cellular subcomponent to another on their way to the extracellular space. These studies are critical for our understanding of how different brain cell types communicate with each other, and more specifically, for characterization of proteins that various brain cell types use for this communication.