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Detecting Released Peptidoglycan Associated Lipoprotein (Pal) from Escherichia coli

Gram‐negative sepsis can result from a bacterial infection, such as Escherichia coli (E. coli), which leads to an over‐exuberant inflammatory response in humans. During E. coli sepsis, a bacterial lipoprotein, Pal (peptidoglycan associated lipoprotein), has been shown to be released from the outer membrane of E. coli and is thought to contribute to an inflammatory response in human cells. In E. coli, Pal has two subpopulations: periplasmic Pal, which is bound to peptidoglycan, and surface‐exposed Pal. A biotinylation technique was used to roughly quantify the amount of each Pal subpopulation that was released from E. coli in vitro. Several different antibiotics were tested for their effect on Pal release; ultracentrifugation was also employed to quantify the amount of Pal released in outer membrane vesicles. Preliminary results suggest that both subpopulations of Pal are capable of release from E. coli, and that the majority of Pal is released from E. coli in outer membrane vesicles.