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Publication
Bifunctional Nitrone-Conjugated Secondary Metabolite Targeting the Ribosome.
Authors Limbrick EM, Graf M, Derewacz DK, Nguyen F, Spraggins JM, Wieland M,
Ynigez-Gutierrez AE, Reisman BJ, Zinshteyn B, McCulloch KM, Iverson TM, Green R,
Wilson DN, Bachmann BO
Submitted By Submitted Externally on 12/3/2021
Status Published
Journal Journal of the American Chemical Society
Year 2020
Date Published 10/1/2020
Volume : Pages 142 : 18369 - 18377
PubMed Reference 32709196
Abstract Many microorganisms possess the capacity for producing multiple antibiotic
secondary metabolites. In a few notable cases, combinations of secondary
metabolites produced by the same organism are used in important combination
therapies for treatment of drug-resistant bacterial infections. However,
examples of conjoined roles of bioactive metabolites produced by the same
organism remain uncommon. During our genetic functional analysis of
oxidase-encoding genes in the everninomicin producer Micromonospora carbonacea
var. aurantiaca, we discovered previously uncharacterized antibiotics
everninomicin N and O, comprised of an everninomicin fragment conjugated to the
macrolide rosamicin via a rare nitrone moiety. These metabolites were determined
to be hydrolysis products of everninomicin P, a nitrone-linked conjugate likely
the result of nonenzymatic condensation of the rosamicin aldehyde and the
octasaccharide everninomicin F, possessing a hydroxylamino sugar moiety.
Rosamicin binds the erythromycin macrolide binding site approximately 60 Å from
the orthosomycin binding site of everninomicins. However, while individual
ribosomal binding sites for each functional half of everninomicin P are too
distant for bidentate binding, ligand displacement studies demonstrated that
everninomicin P competes with rosamicin for ribosomal binding. Chemical
protection studies and structural analysis of everninomicin P revealed that
everninomicin P occupies both the macrolide- and orthosomycin-binding sites on
the 70S ribosome. Moreover, resistance mutations within each binding site were
overcome by the inhibition of the opposite functional antibiotic moiety binding
site. These data together demonstrate a strategy for coupling orthogonal
antibiotic pharmacophores, a surprising tolerance for substantial covalent
modification of each antibiotic, and a potential beneficial strategy to combat
antibiotic resistance.




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