Development of novel affinity reagents to monitor different activation states of Ras GTPases in cells

Type of Award: Catalyst
Date Awarded: June 2012
Award End Date: May 2014
Amount Awarded: $ 200,000.00
PI(s): John P. O'Bryan, PhD, UIC; Shohei Koide, PhD, UChicago;

Abstract: Ras proteins are highly conserved signaling molecules that play critical roles in normal cell growth, and Ras mutations contribute to many forms of human cancer. Thus, molecular mechanisms underlying Ras function has been intensely studied. Ras functions as a molecular switch, cycling between two different states: an active and inactive form. However, we have discovered a third form of Ras that plays an important role in regulating cell signaling. It is generally considered that the third form of Ras exists too transiently in the cell to be functionally important. However, our new discovery suggests that this form is stabilized by interaction with specific proteins also important in cell signaling, thus increasing the lifetime of this form of Ras to a biochemically relevant level that may impact signaling. Our finding is potentially paradigm-shifting in this intensely studied field, and warrants further investigation. A major bottleneck, however, is a lack of suitable research tools. The goal of this proposal is to apply sophisticated protein- engineering technologies to develop biochemical tools that will allow for measurement of the levels of all three forms of Ras. Furthermore, we will use these tools to selectively interfere with the functioning of specific forms of Ras so that we can gain a more complete understanding of their roles in regulating cancer cells.