Indexed on: 29 Jan '19Published on: 28 Jan '19Published in: Journal of Biological Chemistry
Since I started doing scientific research, I’ve been fascinated by the interplay of protein structure and dynamics and how they together mediate protein function. A particular area of interest has been in understanding the mechanistic basis of how lipid signaling enzymes function on membrane surfaces. In this award lecture review I will describe my laboratory’s studies on the structure and dynamics of lipid signaling enzymes on membrane surfaces. This is important as many lipid signaling enzymes are regulated through dynamic regulatory mechanisms that control their enzymatic activity. This review will discuss my continued enthusiasm in using a synergistic application of hydrogen deuterium exchange mass spectrometry (HDX-MS) with other structural biology techniques to probe the mechanistic basis for how membrane localized signaling enzymes are regulated, and how these approaches can be used to understand how they are misregulated in disease. I will discuss specific examples of how we have used HDX-MS to study phosphoinositide kinases, and the protein kinase Akt. An important focus will be on a description of how HDX-MS can be used as a powerful tool to optimize the design of constructs for X-ray crystallography and electron microscopy. The use of a diverse toolbox of biophysical methods has revealed novel insight into the complex and varied regulatory networks that control the function of lipid signaling enzymes, and revealed unique insight into the mechanics of membrane recruitment.