The application for administrative supplement is to request funds to purchase an imaging system to
accelerate the tempo of the research proposed in the parent grant R01 GM084337-01A2, "Small
Molecules that Regulate Proteasome Activity". The overall goal of this study is to identify and
synthesize small molecules derived from natural products that can inhibit or activate the proteasome.
The scope of the overall project is included in the following specific aims proposed to achieve our
goal: (1) To discover new natural product-derived compounds that regulate the proteasome activity.
(2) To develop a next generation of potent proteasome regulators through lead optimization. (3) To
determine the drug binding sites and mechanisms of action. The equipment requested is an
imaging system, such as AlphaInotech FluoChem or Bio-Analyzers. The imaging system is
requested to be used in our lab to efficiently and effectively accomplish the research goals proposed
in the parent grant. This system will use visible light, chemiluminescence or fluorescence to take
visual images of target proteins. The anticipated contribution of the requested imaging system is to
accelerate the tempo of the studies proposed in Specific Aim 3 of the grant. The main objective of
Specific Aim 3 is to identify the target and to elucidate the mechanism of action of the proposed
proteasome inhibitors. As proposed in the research design of the parent grant, a key analysis
proposed to achieve the main objectives is the detection of the proteasome subunit(s) and other
cellular proteins that interact with the proposed proteasome inhibitors. We are currently using an old,
less sensitive Western blot analysis system that detects the targeted proteins by autoradiography.
Due to the low sensitivity and relatively primitive documentation technology of the old system, the
proposed study requires a relatively large quantity of the costly proteasome and cellular proteins, as
well as many more time consuming repetitions to achieve the same goal when compared to an
advanced imaging system. Thus, the requested supplement will greatly speed up the progress of the
proposed study in determining the target and the mechanism of action of the proposed proteasome
inhibitors. In addition to contributing to the progress of the current grant, the requested equipment is
expected be useful for other laboratories in the department because detection of biomolecules, such
as DNA, RNA, and proteins, is one of the most common methodologies used in biomedical science.