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

Innovative and flexible tools for protein purification and molecule binding studies.The goal of this application is to acquire the latest generation of fast protein liquid chromatography system (FPLC) that will be used in combination with a revolutionary isothermal titration calorimetry instrument (ITC). Over the past years, many different research groups have put considerable effort into purifying proteins to investigate their roles, properties, and connections to essential biochemical pathways. However, due to many biophysical limitations and instability of numerous proteins, it has been difficult to purify enough of them to perform their characterization. Thus in order to isolate them in a highly reliable manner, a fast and preparative FPLC system will allow us to overcome these limitations. The requested FPLC has multiple sensors that automatically adjust the optimal pressure along the sample purification. Several important protein properties, including molecular weight, isoelectric point, hydropathicity, hydrophobicity, are automatically taken into consideration during the setup of the system. With an efficient and exquisitely sensitive FPLC system, we will reduce purification time, thereby diminishing overall turnaround time, as well as improving work flow and productivity. Once proteins isolated, our groups subsequently require the use of an isothermal titration calorimetry instrument (MicroCal iTC200) to perform quantitative analyses of the interaction between two or more purified proteins (or ligands) in solution. The MicroCal iTC200 will be an invaluable tool to our research programs as purified proteins (or compounds) that are thought to interact with each other could be measured for their association. The fact that this new generation of iTC requires only very small volumes (uL range) of samples, makes it a method-of-choice for characterizing biomolecular interactions and determining binding affinity between interacting partners that play key regulatory roles in biological pathways.