Dissecting the Molecular Mechanisms of the Histone Acetyltransferase/Cyclic Adenosine Monophosphate Binding Protein Interactome Using Protein-Observed Fluorine NMR

Proteins in biological systems communicate in a type of 'chemical braille', where molecular-scale bumps and grooves of one protein molecule is read by the physical interaction of another protein. Detailed understanding of the molecular-level communications of these protein-protein interactions is necessary to determine the fundamental rules governing both normal and abnormal cellular function. The ability to both characterize and manipulate these interactions at the molecular level has proven challenging. The Pomerantz laboratory at the University of Minnesota works to understanding the chemical details of protein-protein interactions, with the long-term goal of developing ways to stop protein-protein communications that result in harmful cellular functions. To achieve this goal, novel fluorine-based 'probe' protein molecules report on difficult-to-detect protein-protein interactions due to their intrinsic responsiveness in differing chemical environments. Research efforts are integrated with educational activities through course-based undergraduate research experiences (CUREs) designed to increase student success and increase retention in science-technology-engineering-mathematics career paths. Broadened impact of the research project occurs through collaborations with Gustavus Adolphus College, as well as an educational bootcamp that expands undergraduate student exposure to chemical biology research and mentoring by the Pomerantz team.

The Chemistry of Life Process Program in the Division of Chemistry at the NSF is funding Dr. William Pomerantz from the University of Minnesota- Twin Cities for development of fluorine nuclear magnetic resonance spectroscopy (NMR) methods for detecting, quantifying, and defining novel modes of interactions at transcription factor protein-protein interfaces, including those that involve epigenetic regulatory proteins. The long-term research goals of the Pomerantz team are to develop chemical tools to probe the biological roles of transcription factor protein interactions with specific interest in epigenetic gene regulation. This research combines fluorine-19 NMR approaches developed in the Pomerantz lab with additional biophysical methods to characterize native and synthetic ligand binding interactions. This research provides a detailed understanding of the role of aromatic amino acids involved in the protein allosteric network for regulating the KIX protein-protein interaction domain of CREB binding protein (CBP). A recent discovery of the Pomerantz team is the identification of a new protein interaction site on the KIX domain targeted by synthetic small molecules. A key fundamental question to be addressed is the nature of the interaction for other proteins with the newly discovered small-molecule binding site. The Pomerantz team structurally characterizes this site by NMR and uses photo-crosslinking and proteomic methods to evaluate if other proteins occupy this binding site for allosterically regulating KIX function. They evaluate potential domain-domain interactions when studied first as separate domains and ultimately the full-length construct. The research aims are integrated with an educational objective of creating course-based undergraduate research experiences, which result from authentic undergraduate chemistry student research experiences afforded by the study of protein-ligand interactions with fluorine-19 NMR methods.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.