Chemistry
Event Title
An Investigation of the Unique Structural Forms of a Bacterial Enzyme
Document Type
Poster Presentation
Location
Indianapolis, IN
Subject Area
Chemistry
Start Date
11-4-2014 8:30 AM
End Date
11-4-2014 9:30 AM
Sponsor
Jeremy Johnson (Butler University)
Description
Francisella tularensis is a highly infectious bacterium and the causative agent of tularemia. In this project, two different structural forms of FTT258, a serine hydrolase from F. tularensis, were investigated as potential novel drug targets. Specifically, the two distinct structural forms (closed and open) were studied to determine their role in controlling the biological function of FTT258. For these studies, wild type FTT258 was isolated by heterologous expression in Escherichia coli and purification by Ni-metal affinity chromatography. The various protein conformations of FTT258 were then separated by size exclusion chromatography and the relative ratio of the open versus closed forms of FTT258 compared across various conditions. The closed to open form transition was found to be sensitive to time, temperature, and amino acid sequence, suggesting tight cellular control over this transition. The results of these experiments will have direct applications in understanding the regulation of FTT258.
An Investigation of the Unique Structural Forms of a Bacterial Enzyme
Indianapolis, IN
Francisella tularensis is a highly infectious bacterium and the causative agent of tularemia. In this project, two different structural forms of FTT258, a serine hydrolase from F. tularensis, were investigated as potential novel drug targets. Specifically, the two distinct structural forms (closed and open) were studied to determine their role in controlling the biological function of FTT258. For these studies, wild type FTT258 was isolated by heterologous expression in Escherichia coli and purification by Ni-metal affinity chromatography. The various protein conformations of FTT258 were then separated by size exclusion chromatography and the relative ratio of the open versus closed forms of FTT258 compared across various conditions. The closed to open form transition was found to be sensitive to time, temperature, and amino acid sequence, suggesting tight cellular control over this transition. The results of these experiments will have direct applications in understanding the regulation of FTT258.