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Synthesis of a polymyxin derivative for photolabeling studies in the gram‐negative bacterium Escherichia coli
Author(s) -
Meijden Benjamin,
Robinson John A.
Publication year - 2015
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.2736
Subject(s) - polymyxin , escherichia coli , periplasmic space , bacterial outer membrane , chemistry , polymyxin b , gram negative bacteria , bacterial cell structure , lipid bilayer , lipopolysaccharide , biochemistry , bacteria , phospholipid , cell envelope , biophysics , antibiotics , membrane , biology , genetics , gene , endocrinology
The antimicrobial activity of polymyxins against Gram‐negative bacteria has been known for several decades, but the mechanism of action leading to cell death has not been fully explored. A key step after binding of the antibiotic to lipopolysaccharide (LPS) exposed at the cell surface is ‘self‐promoted uptake’ across the outer membrane (OM), in which the antibiotic traverses the asymmetric LPS–phospholipid bilayer before reaching the periplasm and finally targeting and disrupting the bacterial phospholipid inner membrane. The work described here was prompted by the hypothesis that polymyxins might interact with proteins in the OM, as part of their self‐promoted uptake and permeabilizing effects. One way to test this is through photolabeling experiments. We describe the design and synthesis of a photoprobe based upon polymyxin B, containing photoleucine and an N‐acyl group with a terminal alkyne suitable for coupling to a biotin tag using click chemistry. The resulting photoprobe retains potent antimicrobial activity, and in initial photolabeling experiments with Escherichia coli ATCC25922 is shown to photolabel several OM proteins. This photoprobe might be a valuable tool in more detailed studies on the mechanism of action of this family of antibiotics. Copyright © 2015 European Peptide Society and John Wiley & Sons, Ltd.