COBRE: U NEL: CORE B: SPECTROSCOPY

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In order to upgrade the Spectroscopic Facilities, a sequential mixing upgrade for the SX.18MV, Stopped-flow Reaction Analyser, was ordered and received in February 2003. Additionally, an upgrade for the existing Bruker 300E EPR spectrometer was ordered which will greatly enhance the Spectroscopy Facilities. Additionally, a VP-ITC Microcalorimeter was added to the facilities to further enhance the support for biophysical studies. This equipment presents analysis of binding interactions, which is important for the study of redox proteins, most of which first "dock" with their partner, undergo electron transfer and then dissociate. The microcalorimeter will be used extensively by many members of the Redox Biology Center and other faculty at the two institutions to study macromolecule-ligand or substrate and marcromolecule-macromolecule (protein-protein, protein, DNA) interactions. Such data will complement kinetic and thermodynamic data obtained by spectroscopic methods when probes are available and will be unique in systems that do not undergo spectroscopically detectable changes. This facility provides service support for the research in this proposal. Much of the important information about redox reactions and interactions between redox centers is obtained by spectroscopic methods. Electron paramagnetic resonance (EPR) spectroscopy is one of the key instruments used to determine the electronic structure of paramagnetic redox centers. This information complements UV-visible and fluorescence spectra of the macromolecule. Circular dichroism (CD) spectroscopy is used to determine some macroscopic properties of a protein, such as overall conformation between macromolecules and between a macromolecule and a substrate or inhibitor governs the rate of reactions.