Electrophoretic Analysis of Bacterial Cell Wall Peptidoglycan

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Peptidoglycan is a rigid net-like structure that forms the outer wall of bacteria. This molecule gives bacteria their shape and protects them from destruction from outside forces. This "cage" that encloses bacterial cells is constructed by a family of enzymes, the penicillin binding proteins. We know a great deal about the structure and activities of these enzymes themselves, but little is known about how these proteins affect bacterial growth or behavior. One of the major problems in studying the biological functions of peptidoglycan is that current analytical techniques are complex and time-consuming. To address this problem, an electrophoresis procedure is being adapted to analyze the chemical composition of peptidoglycan. This technique, called fluorophore-assisted carbohydrate electrophoresis (FACE), is easier, faster, and more sensitive than currently available techniques. The long term goal of this project is to explain the structure, synthesis, and remodeling of peptidoglycan and to use this information to explain important aspects of bacterial metabolism and physiology. The specific objective of this project is to develop to maturity a simplified, rapid analytical technique that will expand the understanding of peptidoglycan structure and its biological functions. The simplicity and speed of FACE should make it possible to survey the peptidoglycan composition of the bacterial world and lead to new insights about how peptidoglycan structure contributes to bacterial physiology.