|Name||Ms. Ilma Kovac|
|Organization or Institution||University of North Florida|
|Topic||Biochemistry / Chem Bio.|
Development of a screening assay for the Detection of Substrate Specificity Among Bacterial Agmatine Deiminases
Ilma Kovac, Bryan Knuckley
University of North Florida
Agmatine Deiminases (AgDs) are enzymes that are responsible for converting agmatine (a decarboxylated arginine) into N-Carbamoyl Putresciene (NCP). This mechanism is important for the production of ATP (i.e. energy) in a variety of pathogenic bacteria. The objective of this research is to design a new method for detecting substrate specificity among bacterial AgDs. Identifying key differences in the substrate specificity will ultimately lead to more specific inhibitors, therefore producing higher quality antibiotics. To this end, we are characterizing and expressing the AgDs in Streptococcus mutans (SmAgD), Helicobacter pylori (HpAgD), Campylobacter jejuni (CjAgD), Listeria monocytogenes (LmAgD), and Porphyromonas gingivalis (PgAgD). In addition, we are creating a one-bead one compound peptoid library of 100 structures to determine if a specific peptoid sequence reacts with a particular bacterial AgD. The product, a peptoid incorporating NCP, is detected using a chemical probe that turns the beads pink. Comparing the substrate specificity of the various AgDs will provide critical information about the active site, thus leading to a better understanding of inhibitor design.