Phosphatidylinositol 3,4,5–trisphosphate Probe Synthesis and Microarray Protein Binding Studies to Investigate Multivalency and Detect Proteins Upregulated in Cancer

Protein–lipid binding interactions control many processes at the cellular level. Many of these biological events between proteins and lipids are often associated with signaling pathway diseases such as cancer and diabetes. Therefore, increasing our knowledge of protein–lipid binding interactions will provide a better understanding of the signaling pathways of living systems. Pursuing this further, the phosphatidylinositol polyphosphates (PIPs) are an important family of signaling lipids that control key biological processes. Thus, the work described in this thesis was aimed at developing methods to validate these types of lipid–protein binding interactions. The goals of these projects were to synthesize the PIP compound and investigate protein lipid interactions with this compound and others already synthesized using microarray analysis via a pin printer. The results at this juncture in chapter 1 discuss how we were able to achieve successful printing of our target compound as well as determine that binding was occurring. Due to the nature of the microarray we were not able to determine Kd [surface dissociation constant] values for protein–lipid binding interactions due to the fact of not having enough positive data in a singular test event.