Tetracycline does not directly inhibit the function of bacterial elongation factor Tu
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Date
2017
Authors
Gzyl, Katherine E.
Wieden, Hans-Joachim
Journal Title
Journal ISSN
Volume Title
Publisher
Public Library of Science
Abstract
Understanding
the
molecular
mechanism
of antibiotics
that
are
currently
in use
is important
for
the
development
of new
antimicrobials.
The
tetracyclines,
discovered
in the
1940s,
are
a
well-established
class
of antibiotics
that
still
have
a role
in treating
microbial
infections
in
humans.
It is generally
accepted
that
the
main
target
of their
action
is the
ribosome.
The
esti-
mated
affinity
for
tetracycline
binding
to the
ribosome
is relatively
low
compared
to the
actual
potency
of the
drug
in vivo
.
Therefore,
additional
inhibitory
effects
of tetracycline
on
the
translation
machinery
have
been
discussed.
Structural
evidence
suggests
that
tetracycline
inhibits
the
function
of the
essential
bacterial
GTPase
Elongation
Factor
(EF)-Tu
through
interaction
with
the
bound
nucleotide.
Based
on
this,
tetracycline
has
been
predicted
to
impede
the
nucleotide-binding
properties
of EF-Tu.
However,
detailed
kinetic
studies
addressing
the
effect
of tetracycline
on
nucleotide
binding
have
been
prevented
by
the
fluorescence
properties
of the
antibiotic.
Here,
we
report
a fluorescence-bas
ed
kinetic
assay
that
minimizes
the
effect
of tetracycline
autofluorescence,
enabling
the
detailed
kinetic
analysis
of the
nucleotide-bin
ding
properties
of
Escherichia
coli
EF-Tu.
Further-
more,
using
physiologica
lly
relevant
conditions,
we
demonstrate
that
tetracycline
does
not
affect
EF-Tu’s
intrinsic
or ribosome-stimulated
GTPase
activity,
nor
the
stability
of the
EF-
Tu•GTP•Phe-tRNA
Phe
complex.
We
therefore
provide
clear
evidence
that
tetracycline
does
not
directly
impede the function of EF-Tu.
Description
Sherpa Romeo green journal. Open access article. Creative Commons Attribution 4.0 International License (CC BY 4.0) applies.
Keywords
Tetracycline , EF-Tu , Elongation factor , Kinetic analysis , Nucleotide binding , Autofluorescence , GTPase , Inhibit , Antibiotic , Antimicrobial drugs
Citation
Gzyl, K.E., & Wieden, H-J. (2017). Tetracycline does not directly inhibit the function of bacterial elongation factor Tu. PloS ONE, 12(5), e0178523. https://doi.org/10.1371/journal.pone.0178523