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Pyrimidine
Metabolism
in Man.
of Leukemic
I
H.
Lw
By
SMITH,
HE CENTRAL
in protein
tinued
interest
agents
have
precursors
Using
nucleic
in the
the
acids.
These
aspects
of whole
developed
in pyrimidine
which
hemolysates.3
erythrocyte
activities
observed
agents
allow
synthesis
This
and
developed
dthydro-orotic
for
the
deoxyribonucleic
has
and
antineoplastic
pyrimidine
their
greatest
studies
have
colleagues
a con-
or
shown
of purines
clinical
been
in leukemic
have
and
recells.
studied
the
pyrimidines
fractions
of the cells.’
Observations
antineoplastic
agents
on the uptake
and
intact
leukemic
cell.2
Recently
methods
measurements
to be made
report
to
a few
metabolism
his
acids
led
describes
of certain
individual
enzymes
on leukocyte
sonicates
and
the
pattern
of these
enzymatic
cells.
PROCEDURE
of aspartate
carbamytransferase,
dihydro1 ) have
been
described
in detail.3
In
summary,
leukocytes
and erythrocytes
were
separated
by dextran
sedimentation
and
washed
repeatedly.
Aliquots
of cell suspensions
were counted.
Leukocytes
were
destroyed
by sonication
in a Raytheon
10 kc. Oscillator
at maximal
intensity
for 2.5 minutes
and
erythrocytes
by freeze-thaw
hemolysis.
Each
measurement
of an enzyme
activity
involved
the incorporation
of a C14-precursor
of high specific
activity
into the product,
which
was
then
isolated
and analyzed
by the carrier
technic.
Enzyme
assays
were
developed
after
determining
optimal
conditions
of time,
pH,
and
substrate
concentration
( fig. 1).
Aspartate
carbamyltransferase.-C’4-l-aspartate
.015
M; carbamylphosphate
.010
M;
30 mm. incubation;
Tris buffer pH 9.0, 0.20 M.
Dthydro-orotase
and dihydro-orotic
dehydrogenase.-C’4-dihydro-orotate
103
I’sI, 120
mm.
incubation;
Tris
buffer
pH 8.2, 0.25 M.
5-Carboxymethylhydantoinase.-C14-5-carboxymethylhydantoin
10 3 M, 120 mm.
incubation;
Tris buffer
pH 8.2, 0.25 NI.
Following
the addition
of nonisotopic
carrier
and
protein
precipitation
the
product
orotase
methods
acid
precursors
EXPERIMENTAL
The
have
Only
Winzler
isotopic
in leukemic
and
SULLIVAN
respectively,
of nucleic
cells
of
MARGARET
in neoplasia.
A number
of
structural
analogs
of purine
into
the nucleotide
or acid-precipitable
have been made
on the effect of various
utilization
of these
precursors
by the
have been
involved
acids
function,
of leukemia.
of a number
AND
BAKER
ribonucleic
genetic
treatment
quantitative
suspensions
incorporation
of
and
Cells
A.
FArm
in their
metabolism
been
developed
as
of the
on
ROLE
synthesis
effectiveness
ported
JR.,
II. Studies
measurement
dehydrogenase
( fig.
From
the Huntington
Memorial
Laboratories
and the Department
of Medicine,
Massachusetts
General
Hospital
and Harvard
Medical
School,
Boston,
Mass.
This is Publication
No. 984 of the Harvard
Cancer
Commission.
This study
was supported
in part by research
grant
no. A-1606
from
the Institute
of
Arthritis
and Metabolic
Diseases,
and in part by training
grant GRiT
5018 from the National
Cancer
Institute,
U.S.P.H.S.
A preliminary
report
of this work has been published
in abstract
form. (Clinical
Research
7:14, 1959).
Submitted
June 30, 1959; accepted
for publication
Sept. 27, 1959.
360
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PYRIMIDINE
METABOLISM
( carbarnyl
specific
aspartic
activity
IN
acid
or orotic
determined
and
1M
jtNl product
formed
For
comparative
per
10
361
II.
acid ) was isolated
by
total yield calculated
the
carrier
X specific
activity
chroinatographic
by the standard
of the
procedures,
the
carrier
formula:
isolate
=
specific
purposes
leukocytes
or
per
Patients
stuctied.-Normal
clinical
description
of the
MYELOCYTIC
MAN.
activity
all
results
10’
erythrocytes.
are
of the precursor
as mihirnicromoles
expressed
controls
were
obtained
patients
studied
is outlined
Patient
LEUKEMIA.
1. A
42
year
from
below.
old
( mjM
laboratory
woman
with
) synthesized
personnel.
painful
last
therapy
with
Myleran
had
been
completed
3 weeks
prior
to
brief
splenoinegaly
diagnosed
as having
chronic
rnyelocytic
leukemia.
Hemoglobin
10.0 Gm. per
cytes
276,000
per cu.mrn.
with polymorphonuclears
26 per cent,
band
forms
inetamyclocytes
7 per cent, myelocytcs
17 per cent, nucleated
RBC 1 per cent,
I per cent, basophils
2 per cent. There
was a good symptomatic
and hematologic
to x-ray therapy.
Patient
2. A 69 year old man with chronic
myelocytic
leukemia
of 3 years’
been
treated
two times
with splenic
irradiation
and had received
2 courses
His
A
this
was
cent;
leuko46 per cent,
lymphocytes
response
duration
had
of Myleran.
study:
Hemoglobin
8.0 Gm. per cent,
leukocytes,
13,400
per cu.mm.
with
polymorphonuclears
53 per cent,
band forms
16 per cent, rnetamyelocytes
3 per cent, proinyelocytes
1 per cent, rnyeloblasts
1 per cent, lymphocytes
17 per cent,
basophils
5 per cent,
eosinophils
3 per cent,
monocytes 1 per cent. He was considered
to have chronic
myelocytic
leukemia
in partial
remission.
Patient
3. A 69 year old man
with
known
chronic
myelocytic
leukemia
of 2 years’
duration
his
had
been
leukocyte
per
cent,
to
had
rnyelocytes
per
splenic
He
4.
A
cent,
year
basophils
3 per
to
slow
old
Myeleran.
to 361,000
per
promyelocytes
considered
was
79
with
rapidly
per
cent,
was
irradiation
Patient
intermittently
risen
39
7 per
RBC’s
cent.
cleated
Cm.
treated
count
and
woman
have
\Vithin
cu.mm,
9 per
cent,
cent,
month
3 per
myelocytic
prior
to
entry
polymorphonuclears
rnyeloblasts
lymphocytes
chronic
the
with
15
cent.
leukemia
per
24
cent,
nu-
Hemoglobin,
in
relapse.
9.4
Response
incomplete.
entered
the
hospital
with
weakness,
fever,
mild
con-
gestive
failure
and phlebitis:
Hemoglobin,
11.4 Gm. per cent;
leukocytes
260,000
pr cu.
miTi
with
rnyeloblasts
86 per cent, promyelocytes
5 per cent,
myelocytes
4 per cent, polymorphonuclears
3 per cent and lymphocytes
2 per cent. The diagnosis
of acute
myelocytic
leukemia
was
made.
The
patient
died
from
a pulmonary
embolus
within
48 hours.
Patient
5.
bruising
A
43
year
old
woman
entered
the
hospital
because
of
weakness
and
easy
of 3 months
duration:
Hemoglobin,
7.2 Gm. per cent;
leukocytes,
43,000
per en.
mm
with myeloblasts
20 per cent,
promyelocytes
60 per cent,
myelocytes
12 per cent,
polymorphonuclears
6 per cent, and lymphocytes
2 per cent.
The patient
was considered
to have acute or subacute
myclocytic
leukemia.
There was temporary
response
to 6-mercaptopurine
and later to prednisone,
but the patient
died 6 months
later with acute
leukemia
and pneumonia.
1)1 GUGLIELMO
SYNI)ROME.
A 12 year
old boy
presented
with
weakness,
weight
loss,
ecchymoses
and moderate
hepatosplenomegaly.
Hemoglobin
was
7.8 Gm.
per cent and micleated
cells, 8700 per cu.mm
with 50 to 60 per cent being nucleated
erythrocytes.
Reticuloytc count
was
8.8 per cent. Bone
marrow
aspiration
revealed
erythroid
hyperplasia
with
moderate
megaloblastosis.
Over
the
next
8 months
prior to his death
he was treated
with
vitamin
sponse.
B1,
At
nucleated
clears
6-mercaptopurinc,
the
time
cells
16
per
9800
cent,
per
he was
cu.mrn
with
metamyelocytes
forms
cent.
At
nucleated
3 per
autopsy
and
not receiving
cent,
transfusions,
treatment:
erythrocytes
lymphocytes
with
Hemoglobin,
55 per
21
per
only
temporary
3.0
cent,
cent
re-
Gm. per cent,
polyinorphonu-
and
unidentified
there
was extensive
infiltration
of the spleen,
liver,
kidneys,
lymph
nodes
and
marrow
by immature
cells of the erythrocytic
series.
It was
felt by all observers
that his clinical
course,
cytologic
picture
and autopsy
findings
were
consistent
with the Di Guglielmo
syndrome.
MYELOPROLIFERATIVE
DISORDERS.
Patient
1. A 68 year old woman
was followed
in the
blast
5 per
prednisolone
of study
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362
SMITH,
clinic
because
myeloid
of
polycythemia
metaplasia.
At the
of study:
time
2 per
cent,
Patient
ly
140,000
per
cytes 4 per
cent.
year
Leukocyte
myeloid
1 per
man
alkaline
and
was
BAKER
changes
increased
AND
SULL1VAN
suggestive
of
approximately
early
fivefold.
per cent,
leukocytes,
21,300
per
13 per cent,
lymphocytes
5 per
cu.mm
with
cent,
mono-
cent.
with
pulmonary
fibrosis,
phosphatase
emphysema
75
hepatosplenomega-
Hematocrit
per
monocytes
was
and
32 per cent;
leukocytes,
cent,
band
forms
12 per cent, myelo5 per cent,
and nucleated
RBC’s
5
findings:
slightly
elevated.
He
was
considered
to
have
metaplasia.
LYMPHOCYTIC
LEUKEMIA.
enlargement
of
lymph
Biopsy
of
a lymph
Patient
have
2.
A
year
lymphocytic
11.1
cent
and
were
per
revealed
old
girl
entered
months’
lymphoma,
psoriasis
80
cent,
rheumatoid
and
141,000
3 per
hospital
per cent,
monocytes
of
11.8
lymphoblasts
1 per cent.
type.
spondylitis
was
splenomegaly
per
because
Hemoglobin,
lymphoblastic
and
leukocytes,
the
duration:
with
lymphocytes
eosinophils
2 per
lymphadenopathy
cent,
patients
All had
old
several
cu.rnm
cent,
with
with
per
of
malignant
man
polymorphonuclears
Five
studied.
INFECTION.
cu.mm.
Gm.
pruritis
7 per
leukemia
Hemoglobin,
per
and
9000
node
70
1. A 16 year
Patient
nodes
Gm. per cent;
leukocytes,
10 per cent, polymorphonuclears
97
21.4 Gm.
band
forms
cent,
old
splenoinegaly
phosphatase
with the following
hematologic
cu.mni
with polymorphonuclears
cent,
lymphocytes
4 per cent,
presented
per
per
eosinophils
2. A 64
marked
alkaline
Hemoglobin,
78
polyrnorphonuclears
cytes
vera,
Leukocyte
jii.,
cumin.
for
with
known
18
small
to
months:
lymphocytes
cent.
with
pneumonia
a “shift to the
left”
and
leukocytosis
from
in the polymorphonuclear
15,000
to
series.
25,000
RESULTS
(fig.
5-Carboxyinethyihydantoinase
toinase,
which
corresponding
It was
previously
liver.3
as
a qualitative
was
obtained
case
defect
have
in the
enzymatic
the sequence
activities
Di
of
the
it is apparent
tends
a
of
activities
manes
to parallel
with
found
are
orotic
normally
dehydrogenase
absent
from
dihydro-orotase
erythrocytes.
presented
were
only
to its
in Zymobacteriuni
oroticuin.4
leukocytes,
whether
In repeated
erythrocytes
this
studies,
enzyme
no
and
were
present
enzymatic
activity
cells.
studies
of five
presented
been
bar
graphs
patients
above.
of figure
were
sequence
and
three
2, in comparison
increase
the
varied
Di Guglielmo
3. Of
elevated
approximately
whose
obtained
normal
values
were
arbitrarily
of aspartate
car-
From
immaturity
activity
carried
particular
sonicates
in the graphs
dehydrogenase.
syndrome.
in the
patient’s
mature
erythrocytes.
with
patients
activity
parallel.
of
of enzymatic
were
in figure
The
of
in
evidence
on
myelocytic
leukemia,
results
which
were
is maintained
dihydro-orotic
enzymes
cytologic
out
carried
with
The
activities
in leukocytes.
of progressive
increase
syndronie.-Studies
patient
5-carboxymethythydanof carbamylaspartate
in normal
in neoplasia.
that
the
Guglielmo
single
to determine
This
same
dihydro-orotase
bamyltransferase.
eral,
absent
made
leukocytes
summaries
closure
described
leukemia.-Enzymatic
isolated
for these
placed
in
the
been
enzyme
ring
to be
was
in leukemic
presented
are
found
1).-The
reversible
has
An attempt
Myelocytic
the
the
hydantoin,
rat
of
catalyzes
note
erythrocytes,
Aspartate
threefold
the
showing
In gencase
of leukemic
per cell.
out on the
sumcells
erythrocytes
The
enzymatic
is the
finding
of
activities
of dihydro-
an
enzyme
carbamyltransferase
which
is
in comparison
to normal
and
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PYRIMIDINE
METABOLISM
IN
MAN.
363
IL.
0
0
I
ENZ. I - ASPARTATE
CARBAMYLTRANSFERASE
ENZ2 - 5- CARBOXYMETHYLMYDANTOINASE
ENZ.3-DIHYDROOROTASE
ENZ.4-OIHVDROOROTIC
DEHYDROGENASE
0
I
2H(’DPN)
.-
_C__
CH
‘
I
.
..Ct&
N
H
HN
I
#{149}2H
COON
N
H
OA
0
OHO
ENZ4
COOH
ENZ3
#{149}H20 1$20
COOH
I
C
‘CH2
0
0
II
C
HN
HCH
I
H,N-C-O-p-OH
#{149}
NHe
H2N-CH
0.-C
OH
COOH
c.
HCH
HN-CH
!2L
COOH
CAA
COOH
ENZ\HIH
5-CMH
Fig. 1.-Outline
of the enzymatic
CAP,
carbamylphosphate;
1ASP,
5-CMH,
5-carboxymethyihydantoin;
Myeloproliferative
found
as
cells
two
cellular
to
who
the
were
inetaplasia,
respectively,
phatase
determinations,
other
enzymatic
of the
myeloproliferative
figure
111
shift
to the
left,”
of five
such
abnormality
to
cytes.
The
elevation
but
has
leukemia.
the
an
from
immaturity
per
the
dihydro-orotic
In
myeloid
leukocytes
are
cell
being
patients
of
4. Here
and
alkaline
phosto find whether
summarized
are
one
somewhat
of degree.
with
the leticells
per
se, “a
of enzymatic
in figure
been
disorders.5
results
activities
difference
increase
has
vera
between
The
leukocytes
whether
method
control
values
for
with
cell
demonstrated
presented
of
myelocytic
were
made
on
The
findings
are
comparison
be
acid.
in leukemic
and by leukocyte
out in an attempt
and
polymorphonuclear
normal
leukemia.
kemic
with
also
are
difference
phosphatase
polycythemia
grounds
carried
on
orotic
myeloproliferative
enzymatic
carried
out
to determine
leukeinia.-The
only
observations
the
consistent
OA,
cells.
acid;
activity.
The
only
significant
the
dehydrogenase
to about
retwice
value.
concentrate
valid
orotase,
sevenfold).
an
the
have
as in leukemia,
is associated
control
Lyntphocytic
fore
that
studies
was
normal
tends
could
disorders,
studies
were
of infection
sults
of
disorders
acid;
of alkaline
to
on clinical
studies
were
differences
in these
Further
kocytosis
the
leukocytes
4. It is apparent
increased
and
activities
considered
in normal
and leukemic
CAA,
carbamylaspartic
dihydro-orotic
striking
enzymatic
opposed
patients
studied
acid;
DuO,
disorders.-A
the
in
reactions
1-aspartic
normal
similar
marked
for
utilized
for
leukocytes
at
enzyme
leukemia.
lymphocytes
summarized
activities
activities
of aspartate
of
in
leukocytes
are
there-
For comparative
purpos,
however,
from
two patients
with
lymphocytic
in figure
5. It is apparent
that
polymorphonuclear
increase
isolation
of leukocytes
the expense
of lympho-
dihydro-orotic
leukocytes
the
carbamyltransferase
dehydrogenase
lymphocytic
and
in
leu-
dihydro-
( fourfold
to
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
364
5MITH,
JR.,
ASPARTATE
of
CARBAMYLTRANSFERASE
BAKER
enzymatic
of
2500
bamyltransferase,
2000
2000
orotic
dro-orotase
I 500
500
1000
500
1000
500
and
1
4
.
(18)
NORMAL
.
I
-
-
cardihy.
dihvdro.
dehydrogenase
the
leukocytes
patients
with
leukemia
;
activi-
aspartate
2500
.,
SULLIVAN
2.-Comparison
Fig.
the
ties
miM
AND
in
of
five
myelocytic
with
levels
of
activity
found
in normal
polymorphonuclear
leukocytes
(mean
±
S.D.
of 18 determinations).
2
3
4
MYELOCYTIC
LEUKEMIA
DIHYDROOROTASE
miM
1000
1000
800
800
600
600
400
400
2
200
:
(!)
NORMAL
J
34
5
200
MYELOCYTIC
LEUKEMIA
DIHYDROOROTIC
DEHYDROGENASE
mM
300
300
200
200
too
too
U
In
2
()i!
NORMAL
2
3
MYELOCYTIC
4
5
LEUKEMIA
DISCUSSION
The
plastic
in man.
studied
leukocyte
diseases.7
Easily
intact
offers
many
advantages
It is the only
nucleated
isolated
in vitro
as a viable,
without
the
in the study
of metabolic
cell which
can readily
free
floating
cell,
damage
accompanying
the
be
and neoobtained
leukocyte
can
cell preparations
be
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
I’YRIMIDINE
METABOLISM
IN
MAN.
365
H.
miM
muM
50
0
U)
0
a
a
z
100
4
0
Q.
4
0
U4
0
0
50
I 50#{149}
0
4
4
4
4
0
0
}-.cAA
Fig.
3.-Enzymatic
compared
with normal
tions
as in figure
CAA
=
activities
erythrocytes
1. ASP
+
dihydro-orotase,
from
other
and
conveniently
tissues.
cytes
may
of disease
with
certain
destruction
per
on
the
enzyme
cells.
leukemic
corresponding
absent
Its
of the
function
leukemic
has
been
in pyrimidine
myelocytic
the
leukemic
increased
l)articttlarly
In general,
synthesis
the
cell
enzymatic
activity
marked,
increased
being
as
enzymatic
of cells,
as might
immaturity
nonspecific
Leukemic
increase
cells
pathway.#{176}
the
dihydro-orotic
values
for
have
Reduction
normal
leuko.
were
begun
almost
activities
the
in
from
to its
Zymobacteriuni
acid.’
It is
cell tumor.6
has
remained
carhamyltrans-
delwdrogenase
were
mature
polymorphonuclear
of dihydro-orotic
ago.5
absent
acid
with
orotic
the ascites
aspartate
on
studies
years
be
oroticum
of
cells
Such
fifty
was
found
to
1-carbamylaspartic
only
studiedis largely
of blood
investigated.
in Zyniobacteriiim.
and
normal
be determined
the cell population
study
reported
here
variable
cell
can
Finally,
of immaturity
major
demonstrated
The
glvcolvtic
influence
hydantoin,
kocyte.
cells.
of
The
cyclizes
dihydro-orotase
fold above
general
the
cells.
the abnormal
cells
of a number
limitations
in the small
amount
of
5-carboxymethylhydantoinase
This
enzyme,
which
ferase,
several
of
The
was
activities
of
with
DHO±
dehydrogenase.
an organism
isolated
by enrichment
culture
from
many
other
microorganisms,
yeast
and
oroticum,
obscure.
In
myelocyte.
biochemistry
The
number
syndrome
Abbrevia-
carbamyltransferase;
enzymatic
unit
the heterogeneity
and of cell age.
activities
the Di Guglielmo
18 determinations).
aspartate
compared
accompanying
are
of
of
dihydro-orotic
Following
the
enzymatic
CAP±CAA
expressed
“tissue”
obtained
and
in
heterogeneity
of cell type
DHO-0A
the erythrocytes
(mean
±
S.D.
in
DHO±OA
be metabolically
states.
There
concerned
DHO-CAA
increased
len-
dehydrogenase
was
much
as
14 times
elevated
in acute
leukemia.
activity
tended
to parallel
cytologic
evidence
have been
anticipated.
It is to be noted
that
of enzymatic
reduced
of
activities
alkaline
activities
is not
of a number
phosphatase
found
in neoplastic
of enzymes
content
of
in
the
the
leu-
a
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
366
SMITH,
AND
BAKER
JR.,
SULLIVAN
miM
msM
400
a-
0
4
0
(1)
I
a
z
0
4
a:
U-
0
4
200
0
0
a:
0
0
‘5
‘5
(\
4
‘5
.1.)
0
N PNEU.
(5)
UYELPROL
2
(lel
N PNEU. MYELPROL
(18)
(5)
I
2
-.CAA
Fig.
4.-Enzymatic
proliferative
tion
disorders
(pneumonia)
DHO-CAA
activities
in
compared
with
and
N PNEU.MYEL
(IS)
18
the
PROL
I
2
DHO-OA
leukocytes
of
patients
with
five
normal
(5)
controls.
two
patients
the
with
leukocytosis
Abbreviations
are
myelo-
of infec
the
same
as
in
3.
figure
kemic
cell
significance
synthesis
is
of
Winzler
has
isotopic
rates
far
precursors
in excess
consistency
demonstrated
demonstrate
ing
sufficient
to
of the striking
increase
is not apparent
from
these
more
to this
The
specifically
over-all
cell
leukemic
into
normal
leads
course,
acid
to
of
importance.
cell
incorporates
of the
The
in pyrimidine
of whole
cells,
a number
the
acid-precipitable
leukocytes.1’2
The
of
fraction
at
current
studies
individual
enzymes
lead-
synthesis.
increase
unknown.
diagnostic
enzymes
involved
Using
suspensions
in some
of nucleic
which
is, of
the
acids
with
changes
increase
mechanism
neoplastic
that
of nucleic
of those
found
be
of these
studies.
of
This
some
is part
enzymatic
of
the
levels
larger
in
the
problem
of
control
of enzyme
activity
the product
of a series
within
the cell.’0
One such mechanism
for
is that of the negative
feedback
reaction,
in which
of reactions
inhibits
the synthesis
of the proximal
enzymes
own
appropriate
control
involved
for
pyrimidine
E. coli mutants.1’
has
been
resent
genesis
to
the
where
of reaction
in
postulated
the
by enzyme
Di
in
the
formation.
in
added
deletion.
of
types
Dameshek
carhamyltransferase
has
been
neoplasia,
discussed
this
result
E.
with
demonstrated
the
Pardee
on
in that it
leukemia,
This
theory
block
proximal
activity
was
in relationship
occurred
rise
someti’
enzymes.
usually
“pyrimidine
greatest
rep-
is carcino-
in “pyrimidine
the
coli
demonstrated
and
studies
213
formation
of
of enzymatic
In
clearly
including
a hypothetical
it would
increase
in
greatest
increase
has
studies
of Yates
for the current
enzyme
If such
sequence,
dehydrogenase.
of
acquired
syndrome.’4
pyrimidine
This
the
classic
significance
some
results
Guglielmo
dihydroorotic
aspartate
that
metabolic
with
a compensatory
leukemic
cell the
in
its
biosynthesis
This has
rates
In the
found
starvation,”
of
activity.
Re-
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
PYBIMIDINE
METABOLISM
IN
MAN.
367
II.
mpM
miM
2000
600
500
500
a
400#{176}
z
‘5
0
a:
1000
300
U‘5
U
0
:
_I._____
n
I
5OO
N MYEL.
(18) (5)
LYMPH
I
2
5.-Enzymatic
cytic
leukemia.
Although
average
of five studies
cent
studies
blocks
in pernicious
in pyrimidine
enzymatic
that
anemia,
zymatic
this
changes
This
of two
patients
with
values
are compared
18 normal
controls.
there
evidence
and
feedback
in the
LYMPH
I
2
is probably
from
control
leukemic
cell.
the
pattern
as
it
incomplete
is not
The
the
mothis
are
far
results
lymphowith
the
a block
in
have
with
demongenetic
secondary
to B12 deficiency,
with
that
found
in E. coli
indirect
negative
‘5
DHO-OA
in which
synthesis
is consistent
found,
from
N MYEL.
(18) (5)
DHO-CAA
synthesis.15
activities
release
LYMPH
2
activities
in the lymphocytes
of different
cell type,
these
on myelocytic
leukemia
and
later
stages
of pyrimidine
strated
a pattern
which
2OO
nhI1
N MYEL.
(18) (5)
FCAA
Fig.
fl
ia:
‘
the
of increased
is,
would
suggest
of en-
operandi
from
conclusive
in
regard.
In
the
absence
in that
out
mature
erythrocyte
there
of
dihydro-orotic
it does
not contain
protein
be present
is therefore
synthesis.
qualitative
Dihydro-orotic
erythrocytes
difference
in
of nucleic
acid
ing the action
cells.
The
direct
precursors.
of some
by leukemic
direct
studies
function
testing
the
nucleus
agents
have
has
agents
in vitro.
Assay
cells
here
agents
measure
content)
synthesis
on
was
previously
blood
enzyme
this
is found
and
abnormal
erythrocyte.
in the
maturation
sequence
been
developed
methods
individual
as those
enzyme
systems
activities
enzyme
from
fragmented
to
It
as
a
Preunique
as structural
such
the
found
avian
erythrocytes.3
with the appearance
analogs
demonstrated
the feasibility
on the uptake
of nucleic
the
obtained
in
is unique
to carry
nucleated
syndrome
normal
Winzler
of these
of such
reported
of enzyme
peripheral
the
with
pyrimidine
dehydrogenase
the
between
sumably
this enzyme
is lost
in the erythrocyte.
A number
of antineoplastic
allow
in
The
mature
erythrocyte
acid
and therefore
is unable
in the nuclei
of leukocytes
and
of interest
that in the Di Guglielmo
of nucleated
cursors
is a block
dehydrogenase.
ril)onucleic
of studyacid
pre-
described
in
here
leukemic
( presumably
cells.
It
a
is
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
368
SMITH,
understood
trol
that
rates
of
mented
by
rimidine
these
technics
reaction
data
in
of
the
concerning
precursors
necessity
uninjured
the
within
cell.
steady
the
negate
many
Such
a study
state
intact
JR.,
BAKER
AND
factors
SULLIVAN
which
should
concentration
con-
be
of
supple-
individual
pv-
cell.
Sum&ii
1. The
activities
aspartate
genase-were
Neoplastic
orotic
with
much
was
marked
4. The
bacteria,
most
with
in
the
activitate
de
tres
de
orotic-esseva
studiate
con
myelocytic,
leucemia
myeloproliferative,
enzymas
le syndrome
de
Di
con
de
con
esseva
3.
esseva
4.
certe
exhibiva
enzymatic.
Dishydrogenase
patrono
presente
Le
de
Leucocytos
myeloproliferative
simile
dihydro-orotic,
enzyma
absente
del
con
in
some
pyrimidinadihydroab
2
ab
5 patientes
con
un
activitate
de
le evidentia
dihydro-orotic
ab
patientes
sed
minus
es
nucleate
ab
de
leucocytos
con
disordines
patiente
con
omne
le
cytologic
tres
de
cellulas
esseva
infection
marcate
absente
syndrome
hemic
normal
o
le
in
erythrocytos
de
Di
plus
disordine
alterationes
in
del
5-carboxymethylhydantoinase,
esseva
found
e dishydrogenase
erythrocytos
que
in le erythrocytos
hacterios,
previously
hemic
cells.
studiate.
dishydrogenase
augmento
was
syndrome.
lymphocytic,
Le
Le
anormalitate.
similar
erythrocytes,
Guglielmo
circulante
augmentos
parallelismo
frappante
from
showed
in le synthese
leucemia
infection.
Guglielmo
dihydro-
Leukocytes
mature
Di
interessate
sonicatos
tending
of
INTERLINGUA
2. Cellulas
neoplastic
exhibiva
cnzymas,
con
in tendentia
de
immaturitate.
the
pattern.
from
dihydro-orotase,
2
e 5
enzyme
abnormal
IN
aspartato,
in
with
enzymes
disorders
in the
SUMMARIO
1. Le
5 patients
increase
abnormality.
absent
erythrocytes
three
The
striking
enzyme
5-carboxymethylhydantoinase,
was absent
from
normal
and
carbamyltransferase
dehydro-
from
of all
myeloproliferative
alterations
nucleated
synthesis-
2 with
myeloproliferative
from
one patient
with
immaturity.
dehydrogenase,
in the
leukocytes
activities
of
the
or
3. Dihydro-orotic
present
increased
evidence
infection
pyrimidine
dihydro-orotic
studied.
showed
cytologic
less
of circulating
were
dehydrogenase
patients
in
and
2 with
lymphocytic
leukemia,
infection.
The
erythrocytes
cells
the
involved
dihydro-orotase
syndrome
parallel
hut
enzymes
in sonicates
leukemia,
and 5 with
Di Guglielmo
2.
three
studied
myelocytic
disorders
to
of
carbamyltransferase,
le
matur,
Guglielmo.
previemente
incontrate
in
e anormal.
REFERENCES
1.
Winzler,
nucleic
human
Etiology,
ment.
1957.
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agents
and
acid
metabolism
of isolated
leukocytes.
In The Leukemias:
Pathophysiology,
and TreatNew
York,
Academic
Press,
R.
2.
-:
l)ifferences
lism
in
between
human
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L.
Pyrimidine
nucleic
normal
leukocytes.
75:37,
1958.
H., Jr., and
metabolism
acid
iiietabo-
and
Ann.
letikemic
New
Baker,
in man.
York
F. A.:
I. The
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369
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acid.
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1958.
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A. B.: The control
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1959,
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4.
Lieberman,
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A.:
Enzymatic
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orotic
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II. Dihydroorotic
acid,
ureidosuccinic
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and
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W. S.:
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N.:
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M.,
and
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vol.
Ed.:
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In Tocantins,
in Hematology.
Stratton,
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1956,
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wechsels
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Blutzellen).
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zur
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weissen
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R.
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E. E.:
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phomas,
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L.
Pyrimidine
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J.Nat.
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Leuk#{228}mic
(Zugleich
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leu-
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ane-
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1960 15: 360-369
Pyrimidine Metabolism in Man. II. Studies of Leukemic Cells
LLOYD H. SMITH, JR., FAITH A. BAKER and MARGARET SULLIVAN
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