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Effect of penicillin on the uptake of amino acids in bacteria
Vol. 1, No. 1
BIOCHEMICAL
EFFECT
OF
AND BIOPHYSICAL
PENICILLIN
ON IN
THE
In
the
course
inhibition
of
arginine
(Gorini
begun
with
exponentially by
arginine
which
converts of
hydrolysate increased A defect and
to
cells
medium
the
level
the
Simon,
(Gale
acid
and
therefore the
presence
arginine
*
so
to
A
Taylor,
1947).
measured
in
of
penicillin.
prevent
was 1958)
percent
sucrose of
penicillin
repression
ornithine
to
ordinarily
transcarbamylase,
About
60
a yeast
minutes
after
the
extract-casein
the
enzyme
began
the
release
of
to
rise
and
minutes.
the
been
formation
addition
in
of
(Sistrom,
the
of
mechanism that
had
the
feed-back
biosynthesis
enzyme
20
abolished
for
gradient.
glutamic
that
of
30
concentrating 1959)
of
growing
explanation
concentration of
within
of
of
the
(O&S)
citrulline.
penicillin
possible in
to
ACIDS
Medicine
mechanism
study
synthesis
of
in
presence
noted
the
ornithine
ZO-fold
a
culture
on
AMINO
York
the
spheres
the
was
growing
exerted
addition
It
New
on
sensitive
1956).
Microbiology College
16,
1957)
in
OF
Maas*
(repression)
Maas,
penicillin
(Lederberg,
York
formation and
of
investigations
osmotically
prepared
an
of
enzyme
using
K.
Department York University New
UPTAKE
July 1959
BACTERIA
Werner
New
RESEARCH COMMUNICATIONS
cell
for
noted The
cultures
arginine
cannot
similar
of
by
growing
20
Chloramphenicol
incorporation
The author is a Senior Service, SF-129. The work the National Science Foundation Health Service. The excellent Kolodny is acknowledged.
of
Gale
and
amino
the
percent
acid
sucrose added into
was in
before protein,
Research Fellow, U. S. Public was supported by grants No. and RG-6048 from the U. technical assistance of Mrs.
13
uptake
Taylor
arginine
was the
enough
on
radioactive in
a Maas
a large
penicillin
previously of
is
(Schwartz,
maintain
effect
uptake
repression
Health G-3344 S. Public Roslyn
from
Vol. 1, No. 1
As
BIOCHEMICAL
shown
in
to
concentrate
cell penicillin
Table
is
with
penicillin.
the
action
after
added
of
penicillin of
arginine,
Under
these the
is
but
pronounced
conditions
30
effect
after slight
protected
chloramphenicol at
240
percent
60
minutes
minutes
sucrose
from
decreases
chloramphenicol
minutes being
the
effect
for
a number
see
Schwartz,
similar the
of of
penicillin
other
uptake
and in
of
and
lytic
Experiments is
necessary
These or
forms
acid
action
Hoare
penicillin,
in to
a few The
l/3
in
presence the of
as
a loss
concentrating
the of
in
Bacillus
a
have
indicated
that
inhibit
uptake
to
with or
of
by
omitting
in
the
reached
in
cells.
OSS
behave
differently
uptake
is
whole
concentrating penicillin’s
not
up
the
to
cell
wall
mechanism capacity
acids. 1950)
auxotroph
Even
(tested
amino
diaminopimelic
is
of
of
cell
lysozyme(Sistrom,
arginine
penicillin
intact
a DAP-requiring
OSS
arginine
an
of
concentrated
only
This
is
from
10,000
per
resulted
in
amino
interfere
of cells.
further units
has
one
intact
depressed
for to
absence
acids
with
this
for
the
ml). an as
mechanism.
Apparently inhibition
result
organism
1957).
integrity
to
gram-positive
of
which of
of
altering
type level
level
impairment well
Work,
the
lowered
Thus,
and
found
D-serine,
inhibited
1956) medium
was
and
penicillin
either
the
glycine
Furthermore,
penicillin
each
instances
the
OSS
mechanisms
I).
prepared
from
(Meadow,
l/4
with
(Lederberg,
(DAP)
to
(Table
were
penicillin
1959)
the
KM
for
concentrating (lysine,
Simon,
in
strain
the
acids
uptake.
arginine
megaterium
wall
amino
Maas
decrease
on
of
protein uptake
by
synthesis penicillin.
is
also
In
various
14
required amino
the
when
of
after
(20
of
and
addition
added
the
are
maximal
the
slow,
cells
is
ability
chloramphenicol
between
is
the
inhibition with
penicillin
inhibits
July 1959
penicillin. The
in
The
intervals
When
I).
markedly
concomitantly
chloramphenicol) of
penicillin
time
addition
(Table
I,
arginine.
increasing
and
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
acid
auxotrophs
37’
Chloramphenicol in an arginine-free,
strain
(final
KM
concentration enriched medium
*
Uptake
intracellular
of
C’
’
arginine
concentrations
measured
of
arginine
240
minutes
were
t
488
1400
after
calculated
its
0.5
exponentially percent
(1790)*
bacteria
from
addition.
the
specific
activity
of
the
preparation.
1000 units/ml) was added at times labelled, was added (final concentration of the cultures were then collected (Atkinson and McFadden, 1956). The
cells 1958)
985
1800
3260
penicillin
weight
935
growing with
no
wet
2100
(ca5)*
I pg/g
403
penicillin
Uptake
200 pg/ml) was added to (AF) (Gorini and Maas,
0
0
and
addition
lactate as carbon source. Penicillin (final concentration indicated, Five minutes later, C’ 4- arginine, randomly 20pg/ml) and incubation continued for 60 minutes. Aliquots on membrane filters and their radioactivities determined
at
megaterium,
30
,I
B.
10
W
I
minutes
between
chloramphenicol
penicillin,
of
Interval
II
strain
coli,
E.
Organism
TABLE
Vol. 1, No. 1
BIOCHEMICAL
IlysineJ
isoleucine,
concentration growth
of
will
of
restore
even effect
the
the
inhibition
It
such
1957)the
observed
in
as
miy
be
inhibition
the
of
one
the
a
there
the
is
early
no
of
the
sets
(Park
formation
lysis,
manifestations
which
synthesis
small
these
Thus
action
wall
required
penicillin
Under
of
trigger
cell
in
of
but
the of
chloramphenicol. is
the
of
presence
July 1959
the
exhaustion
occurs,
or
prevent
exhaustion
uptake.
here
unbalance
cell
in
mass
sucrose
not
after
arginine
described
antibiotic.
changes,
addition
of
of
after
acid
increase
penicillin
finally
the amino
absence
does
arginine
required the
of
and
radioactive
a small in
penicillin
However, the
conditions
of
arginine)
factor.
amount
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
off
further
and
Strominger,
repressible
enzymes,
lysis.
BIBLIOGRAPHY Atkinson,
D.E.,
Gale,
E. F ., and
Gorini,
L.
Gorini,
L. W. D.
J.
Meadow,
P.,
Schwartz,
Sistrom,
Received
581
Gen.
“The W.K., B. Glass 1958.
Natl.
Acad.
D. S.
and
Acta
Chemical
Basis
editions,
Sci.
J.L.
E.,
of
The
Johns
-42,
574
U.S.
Biochem.
Science
and
- 71, 1,
Biophys.
Work,
W.K.,
Bacterial.
Microbial.
Biochim.
Strominger, Maas,
J.
125,
Simon,
E.
Biochim.
21,
Biophys.
Acta
1959
16
-29,
(1947).
-25,
208
(1957).
Hopkins
University
(1956). - 66,
270
(1957)
(1957).
Biochim.
579
(1956).
314
J. 99
123
Development”,
(B59).
W.R.,
June
and
Hoare,
J.H., --32,
W.K.,
Proc.
and
J.T.,
B.A.,
E. S. , J.
Maas,
and Maas, McElroy Baltimore,
Lederberg,
McFadden,
Taylor,
and
Press,
Park,
and
(1958).
Biophys.
Acta
BIOCHEMICAL
EFFECT
OF
AND BIOPHYSICAL
PENICILLIN
ON IN
THE
In
the
course
inhibition
of
arginine
(Gorini
begun
with
exponentially by
arginine
which
converts of
hydrolysate increased A defect and
to
cells
medium
the
level
the
Simon,
(Gale
acid
and
therefore the
presence
arginine
*
so
to
A
Taylor,
1947).
measured
in
of
penicillin.
prevent
was 1958)
percent
sucrose of
penicillin
repression
ornithine
to
ordinarily
transcarbamylase,
About
60
a yeast
minutes
after
the
extract-casein
the
enzyme
began
the
release
of
to
rise
and
minutes.
the
been
formation
addition
in
of
(Sistrom,
the
of
mechanism that
had
the
feed-back
biosynthesis
enzyme
20
abolished
for
gradient.
glutamic
that
of
30
concentrating 1959)
of
growing
explanation
concentration of
within
of
of
the
(O&S)
citrulline.
penicillin
possible in
to
ACIDS
Medicine
mechanism
study
synthesis
of
in
presence
noted
the
ornithine
ZO-fold
a
culture
on
AMINO
York
the
spheres
the
was
growing
exerted
addition
It
New
on
sensitive
1956).
Microbiology College
16,
1957)
in
OF
Maas*
(repression)
Maas,
penicillin
(Lederberg,
York
formation and
of
investigations
osmotically
prepared
an
of
enzyme
using
K.
Department York University New
UPTAKE
July 1959
BACTERIA
Werner
New
RESEARCH COMMUNICATIONS
cell
for
noted The
cultures
arginine
cannot
similar
of
by
growing
20
Chloramphenicol
incorporation
The author is a Senior Service, SF-129. The work the National Science Foundation Health Service. The excellent Kolodny is acknowledged.
of
Gale
and
amino
the
percent
acid
sucrose added into
was in
before protein,
Research Fellow, U. S. Public was supported by grants No. and RG-6048 from the U. technical assistance of Mrs.
13
uptake
Taylor
arginine
was the
enough
on
radioactive in
a Maas
a large
penicillin
previously of
is
(Schwartz,
maintain
effect
uptake
repression
Health G-3344 S. Public Roslyn
from
Vol. 1, No. 1
As
BIOCHEMICAL
shown
in
to
concentrate
cell penicillin
Table
is
with
penicillin.
the
action
after
added
of
penicillin of
arginine,
Under
these the
is
but
pronounced
conditions
30
effect
after slight
protected
chloramphenicol at
240
percent
60
minutes
minutes
sucrose
from
decreases
chloramphenicol
minutes being
the
effect
for
a number
see
Schwartz,
similar the
of of
penicillin
other
uptake
and in
of
and
lytic
Experiments is
necessary
These or
forms
acid
action
Hoare
penicillin,
in to
a few The
l/3
in
presence the of
as
a loss
concentrating
the of
in
Bacillus
a
have
indicated
that
inhibit
uptake
to
with or
of
by
omitting
in
the
reached
in
cells.
OSS
behave
differently
uptake
is
whole
concentrating penicillin’s
not
up
the
to
cell
wall
mechanism capacity
acids. 1950)
auxotroph
Even
(tested
amino
diaminopimelic
is
of
of
cell
lysozyme(Sistrom,
arginine
penicillin
intact
a DAP-requiring
OSS
arginine
an
of
concentrated
only
This
is
from
10,000
per
resulted
in
amino
interfere
of cells.
further units
has
one
intact
depressed
for to
absence
acids
with
this
for
the
ml). an as
mechanism.
Apparently inhibition
result
organism
1957).
integrity
to
gram-positive
of
which of
of
altering
type level
level
impairment well
Work,
the
lowered
Thus,
and
found
D-serine,
inhibited
1956) medium
was
and
penicillin
either
the
glycine
Furthermore,
penicillin
each
instances
the
OSS
mechanisms
I).
prepared
from
(Meadow,
l/4
with
(Lederberg,
(DAP)
to
(Table
were
penicillin
1959)
the
KM
for
concentrating (lysine,
Simon,
in
strain
the
acids
uptake.
arginine
megaterium
wall
amino
Maas
decrease
on
of
protein uptake
by
synthesis penicillin.
is
also
In
various
14
required amino
the
when
of
after
(20
of
and
addition
added
the
are
maximal
the
slow,
cells
is
ability
chloramphenicol
between
is
the
inhibition with
penicillin
inhibits
July 1959
penicillin. The
in
The
intervals
When
I).
markedly
concomitantly
chloramphenicol) of
penicillin
time
addition
(Table
I,
arginine.
increasing
and
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
acid
auxotrophs
37’
Chloramphenicol in an arginine-free,
strain
(final
KM
concentration enriched medium
*
Uptake
intracellular
of
C’
’
arginine
concentrations
measured
of
arginine
240
minutes
were
t
488
1400
after
calculated
its
0.5
exponentially percent
(1790)*
bacteria
from
addition.
the
specific
activity
of
the
preparation.
1000 units/ml) was added at times labelled, was added (final concentration of the cultures were then collected (Atkinson and McFadden, 1956). The
cells 1958)
985
1800
3260
penicillin
weight
935
growing with
no
wet
2100
(ca5)*
I pg/g
403
penicillin
Uptake
200 pg/ml) was added to (AF) (Gorini and Maas,
0
0
and
addition
lactate as carbon source. Penicillin (final concentration indicated, Five minutes later, C’ 4- arginine, randomly 20pg/ml) and incubation continued for 60 minutes. Aliquots on membrane filters and their radioactivities determined
at
megaterium,
30
,I
B.
10
W
I
minutes
between
chloramphenicol
penicillin,
of
Interval
II
strain
coli,
E.
Organism
TABLE
Vol. 1, No. 1
BIOCHEMICAL
IlysineJ
isoleucine,
concentration growth
of
will
of
restore
even effect
the
the
inhibition
It
such
1957)the
observed
in
as
miy
be
inhibition
the
of
one
the
a
there
the
is
early
no
of
the
sets
(Park
formation
lysis,
manifestations
which
synthesis
small
these
Thus
action
wall
required
penicillin
Under
of
trigger
cell
in
of
but
the of
chloramphenicol. is
the
of
presence
July 1959
the
exhaustion
occurs,
or
prevent
exhaustion
uptake.
here
unbalance
cell
in
mass
sucrose
not
after
arginine
described
antibiotic.
changes,
addition
of
of
after
acid
increase
penicillin
finally
the amino
absence
does
arginine
required the
of
and
radioactive
a small in
penicillin
However, the
conditions
of
arginine)
factor.
amount
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
off
further
and
Strominger,
repressible
enzymes,
lysis.
BIBLIOGRAPHY Atkinson,
D.E.,
Gale,
E. F ., and
Gorini,
L.
Gorini,
L. W. D.
J.
Meadow,
P.,
Schwartz,
Sistrom,
Received
581
Gen.
“The W.K., B. Glass 1958.
Natl.
Acad.
D. S.
and
Acta
Chemical
Basis
editions,
Sci.
J.L.
E.,
of
The
Johns
-42,
574
U.S.
Biochem.
Science
and
- 71, 1,
Biophys.
Work,
W.K.,
Bacterial.
Microbial.
Biochim.
Strominger, Maas,
J.
125,
Simon,
E.
Biochim.
21,
Biophys.
Acta
1959
16
-29,
(1947).
-25,
208
(1957).
Hopkins
University
(1956). - 66,
270
(1957)
(1957).
Biochim.
579
(1956).
314
J. 99
123
Development”,
(B59).
W.R.,
June
and
Hoare,
J.H., --32,
W.K.,
Proc.
and
J.T.,
B.A.,
E. S. , J.
Maas,
and Maas, McElroy Baltimore,
Lederberg,
McFadden,
Taylor,
and
Press,
Park,
and
(1958).
Biophys.
Acta