[CANCER RESEARCH 27, 117-123,January 1967]
Developmental Changes of Hepatic Catalase in the Rat1
GRANT W. PATTON2 AND EDWIN T. NISHIMURA3
Department of Pathology, Northwestern University Medical School, Chicago, Illinois,
fornia School of Medicine, Los Angeles, California
Summary
Immunoelectrophoretic analysis of 25-day-old newborn and
adult rat livers verified the presence of a 6-arc pattern of catalase
observed earlier for purified rat hepatic catalase and for liver
homogenates. Application of the immunoelectrophoretic method
to fetal and newborn liver homogenates revealed a gradual shift
from predominantly anodic arcs present in the fetal and early
newborn period to largely cathodic arcs present in the 14-day
newborn animals, followed by a full complement of catalase sub
components present in 25-day-old and adult livers. The pattern
due to erythrocyte catalase subcomponents present in liver
homogenates could be readily differentiated from the hepatic
catalase pattern.
Quantitative assay of all liver homogenates showed a 4- to 5fold increase in catalase activity during the 16- to 20-day fetal
period and correlated with an increased concentration of a single
subcomponent. Little alteration in enzyme activity occurred
during the perinatal period. The mean value of hepatic catalase
activities at 14 days was lower than the early newborn period
and was accompanied by a shift in prominence of cathodic sub
components associated with the absence of 2 anodic arcs seen
in younger fetuses. The livers of 25-day newborn animals
demonstrated adult levels of catalase activity and an adult-type
pattern.
Introduction
It has been suggested recently that plant and animal catalases
are structurally heterogeneous molecules composed of molecular
subunits (2, 12, 20). Numerous studies of purified crystalline
hepatic catalase have demonstrated a separation of the enzyme
into smaller fragments (1, 15, 16, 19, 20); however, only a few
observers found the smaller molecular weight components to be
enzymatically active (4, 11, 12, 20). Studies by Samejima (15,
16) suggest that BHC4 is composed of 6 subunits; others (17,
1 Supported in part by TJSPHS Grant No. CA 08596-01 and a
grant from the California Institute for Cancer Research.
2 Present address: Harvard Medical School, Boston Lying-in
Hospital, Boston, Mass.
1 Present address: Department
of Pathology, University of
Hawaii School of Medicine, Honolulu, Hawaii.
4 The following abbreviations
are used: RHC, rat hepatic
catalase; REC, rat erythrocyte
catalase; BHC, bovine liver
catalase; BEC, bovine erythrocyte catalase; HHC, human liver
catalase; HEC, human erythrocyte catalase; AbRHC and AbREC,
rabbit antisera to RHC and REC, respectively.
Received May 16, 1966; accepted August 18, 1966.
and Department of Pathology, University of Cali
20) have expressed the view that bovine catalase may be com
posed of only 4 subunits. Nishimura et al. has shown that 6
immunoelectrophoretic components of RHC may be arranged
in a manner reminiscent of the pattern for lactic dehydrogenase
(12).
A number of investigators have demonstrated immunologie
differences in catalases from various species (3, 5, 8, 12); of
greater significance, however, are the electrophoretic and im
munologie variations of catalases from different organs of the
same animals (13). The amino acid compositions of equine liver
catalase and equine erythrocyte catalase have been shown to be
different (14), and each has revealed some differences in the
absorption spectra (3). The serologie differences in the 2 forms of
bovine catalase, moreover, have been repeatedly demonstrated
(6, 9, 14).
An immunoelectrophoretic survey of catalase isozyme patterns
in adult rat organs utilizing hepatic and erythrocyte catalase
antisera was recently reported by Nishimura et al. (13). The
enzyme pattern of kidney was observed to be similar to the
hepatic form, but striking differences in the immunoelectro
phoretic patterns were noted for spleen, heart, gonads, skeletal
muscle, and other tissues (13).
Evidence for differences in the intracellular forms of hepatic
catalase also have been demonstrated. Brown found 2 forms of
BHC (4), and, in an analysis of the subcellular fraction of rat
liver, Higashi and Peters (8) described 2 distinct forms of mitochondrial catalase.
Little information has appeared in the literature with reference
to the development of hepatic catalase in the maturing fetus and
the newborn. Greenstein in 1942 (7) observed that fetal livers of
the rat possessed only yV of the amount of catalase found in adult
livers. In a more comprehensive study Stevens (18) followed the
progress of catalase development in the livers and kidneys of rats
during the growth of the fetus and the newborn. These studies,
however, have been limited to the quantitative assessment of
enzyme activities and compared only the catalytic actions of
tissues of one stage of development to those of another. Recent
studies have shown that liver catalase of the adult rat consists of
6 distinct immunoelectrophoretic subcomixments (12). Since the
maturing liver exhibits histologie, and undoubtedly physiologic,
transitions from the fetal to a more uniformly organized adult
feature, a series of experiments was conducted to correlate the
variations in enzymatic activities with the immunoelectro
phoretic patterns of liver catalase during the maturation of the
rat fetus and the newborn.
JANUARY 1967
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117
Grant W. Patton and Edwin T. Nishimura
AB RHC
AB RHC
H6
H6
CHART 1. Typical immunoelectrophoretic
pattern of purified rat liver catalase reacting with rabbit anti-rat hepatic catalase serum
(AbRHC) is illustrated in the figure to the left. The other diagram, to the right, depicts the pattern observed for crude, adult liver
homogenate sample.
AB REG
AB RHC
HI
\
E3
—E 6
PURE
RHC
CHART 2. Fusion of identity is represented
and E-6. See text for discussion.
Materials
between Arc H-4
and Methods
Anticatalase sera against adult rat hepatic or erythrocyte
catalase antigen were developed in New Zealand rabbits as
described earlier (12, 13). Purified RHC with a Katf (for defini
118
tion see Ref. 21) of 69,000 was used for the induction of AbRHC.
Highly purified REC, Katf 120,700, was utilized for the induction
of AbREC. Control sera were obtained from 2 rabbits which
received injections of equivalent amounts of Freund's adjuvant
alone for the same i>eriod of time.
Sprague-Dawley rats of the Holtzman strain were bred and
surgically delivered through an abdominal incision under ether
anesthesia at defined intervals after conception. The viable
fetuses were bled, and their livers were removed, washed in
isotonic saline, and quickly frozen with Dry Ice. Additional
tissue samples from each litter were separated, fixed in formalin,
and embedded in paraffin, and the sections were stained with
hematoxylin and eosin to verify the fetal age. Further verifica
tion of fetal age was obtained by the correlation of morphologic
development and crown-rump length which were compared
against the accepted standard. Because of minute quantities of
tissues, the livers from all fetal and early-newborn litter mates
were pooled and assayed in duplicates. In the late-newborn and
adult animals, the hepatic tissue from individual animals was
analyzed separately. Livers from healthy adult rats of both sexes
weighing about 250 gm were used as controls.
A modification of the classic method of von Euler and Josephson previously described (13) was employed for the catalase
assay of diluted homogenates. This procedure utilized 5 ml of
0.01 M H20i in 0.01 M phosphate buffer at pH 6.8 as substrate,
at 37°C,to which 1 ml of diluted liver homogenate sample was
added by means of a blow-out pipet. The reaction was timed
for 15, 30, 45, and 60 sec and terminated by addition of 2 ml of
2 N HzSO.!.The excess of H20s in each test was back-titrated with
a standardized 0.01 x KMnOi to a faint pink end point. The Ist
order reaction constant K, was calculated from the equation
Kt =
2.303
X log.o
ml KMnO4 blank
ml KMnU4 unknown
A plot of A'Õ
values permitted extrapolation to A'0for the enzyme
CANCER
RESEARCH
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VOL. 27
Changes of Hepatic Catalane
at 0 time. Catalase activity was expressed in relation to nitrogen
content (K/N). The nitrogen content of homogenate samples was
determined by the micro-Kjeldahl technic.
The methods of immunoelectrophoretic analyses of purified
rat catalase and tissue homogenates have been described earlier
(13) and utilized lonagar No. 2 buffered at pH 8.6 with Hirsch
feld modification of Laurell's barbital buffer containing calcium
lactate. A 2-mm layer of 1.15% agar was applied on 5- x 7.5-cm
glass slides. Purified catalase and homogenate samples of 0.025
ml each were applied to centrally located wells, and electrophoresis was carried out at 3°-5°C
with 23 volts/cm of slide
width for 2 hr. About 0.2 ml of antiserum was introduced into
each of the longitudinal troughs located on either side of the well
and diffusion carried out for 48-72 hr at 8°C.
Results
Immunoelectrophoretic
Patterns of Adult Liver Homogenate
Data from adult liver homogenate samples are presented in
Chart 1. The pattern corresponds closely to that of purified adult
rat hepatic catalase when it is reacted against AbRHC and
AbREC (Chart 2). The 6-arc pattern of purified liver catalase
developed with homologous antiserum is contrasted with 2
antigenically similar arcs produced by the antiserum AbREC.
One is faint and uncertain and represented with a broken line.
Arcs H-4 and E-6 show fusion of identity at the cathodic ends;
it is not absolutely certain whether the reacting liver subcom
ponent is Arc H-4 or H-5 as noted earlier (13). The hepatic and
erythrocyte subcomponents are identified by immunoelectro
phoretic precipitin arcs; they have been labeled in sequence from
the anodic pole as shown in Charts 1 and 4.
For controls of immunoelectrophoretic tests, purified hepatic
catalase and adult liver homogenates were reacted against the
control antisera developed in rabbits injected with Freund's
adjuvant alone. Such tests always gave a negative precipitin
reaction. Similar controls were performed on representative
samples of fetal and newborn liver homogenates discussed below.
Precipitin arcs were not observed in any fetal or newborn homog
enates tested with control serum.
Fetal and Newborn Patterns with AbRHC
Chart 3 is a diagrammatic representation of immunoelectro
phoretic variations observed when liver homogenates from fetal
and newborn rats were reacted with AbRHC. The 16-day fetal
liver demonstrates 4 subcomponents corresponding to Arcs H-l,
H-2, H-3, and H-6 of the adult form. In the 19-day fetus, Arc
H-6, faint at 16 days, becomes better defined, and this 4-arc
pattern persisted through Days 20 and 21 of fetal development.
A unique anodic arc, which did not corres[x)nd to any adult
form, appeared in the 22-day fetus and persisted though the 24hr newborn period. Immunoelectrophoretic patterns of the 2-hr,
12-hr, and 24-hr newborn rat liver catalase samples were iden
tical. The 14-day newborn liver demonstrated a rather re
markable change in enzyme subcomponents. Anodic Arcs H-2
and H-3, seen in earlier samples, were no longer present, and
cathodic Arcs H-4 and H-5 made their appearance at this time.
This 4-arc pattern was consistent in all animals examined at this
age. Finally, the 25-day newborn animals demonstrated an adult
pattern with the full complement of 6 arcs, including Arcs H-2
and H-3, which had been temporarily absent shortly prior to
birth.
In summary, the anodic subcomponents of hepatic catalase
appear to develop early and are accompanied by a single cathodic
arc until a period immediately preceding parturition, when an
anodic arc, unique for this stage of development, appears. At
14 days, newborn liver catalase contains predominantly cathodic
subcomponents which are accompanied by a single anodic arc.
Finally, at 25 days after birth the pattern appears similar to that
of the adult form.
Fetal and Newborn Patterns with AbRHC and AbREC
The possibility that portions of the immunoelectrophoretic
patterns presented above might be due to erythrocyte catalase
contamination of saline-washed livers was considered in the
following 2 experiments. In the 1st experiment, purified adult
rat erythrocyte catalase was reacted against AbRHC to ascertain
the antigenic subcomponents of the erythrocyte enzyme which
might cross-react with AbRHC and thus confuse the interpreta-
DIAGRAMS OF IMMUNOELECTROPHORETIC
PATTERNSOF FETAL, NEWBORN
AND ADULT RAT LIVER HOMOGENATESREACTEDWITH Ab RHC
FETAL
FETUS
16 d
CHART 3. A comparison
stages.
JANUARY
of variations
FETUS
19 d
NEWBORN
FETUS
20 d
FETUS
22 d
in immunoelectrophoretic
ADULT
ADULT
NEWBORN NEWBORN
(& 25 DAY
24 hr
14 d
N.B.)
patterns
of liver homogenates
during fetal, newborn,
and adult
1967
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119
Grant W. Patton and Edwin T. Nishimura
tion of such homogenate studies. Chart 4, representing the
reaction of purified erythrocyte catalase against AbREC and
AbRHC, demonstrates a single cross-reacting arc tentatively
identified as H-4 and E-6. The arc corresponding to E-3 and
illustrated in broken line (Chart 2) when purified liver catalase
was reacted against AbREC was not seen, and this discrepancy
is, at present, unexplained. The single cross-reacting arc of
purified REC also was seen when RBC homogenates were used
instead of purified erythrocyte catalase.
In the 2nd experiment each fetal and newborn liver homog
enate was reacted with AbRHC and AbREC on the same agar
plate. The results are presented in Chart 5. Fetal liver homogAB REC
AB RHC
enates at age 16 days have 4 arcs when reacted against AbREC,
which arc unrelated to arcs developed against AbRHC, and at
age 19 days reveal only 2 precipitili arcs against AbREC. Similar
homogenate preparations of 20-, 21-, and 22-day fetuses also
revealed 2 arcs which are similar to the pattern seen in adult
liver homogenate when reacted with AbREC; a 3rd short arc
near the well (origin) was also observed in some of the prepara
tions of this fetal age group. In these samples, hepatic Arc H-6
intersected the long erythrocyte arc resulting in a spur formation
(see Pattern C, Chart 5) when the reaction was permitted to
"overdevelop." The 2-, 12-, and 24-hr newborn liver samples
demonstrated patterns which were essentially identical with the
pattern of the 22-day fetus (D and E, Chart 5). A sample from
22-day fetus revealed antigenic similarity between Arc H-l and
an erythrocyte arc, possibly E-3. In the 14-day newborn rat
liver (F, Chart 5), an arc migrating to the anodic pole was
observed and may represent a subcomponent similar to E-l or
E-2. In the 25-day newborn sample, the liver-homogenate
pattern of catalase subcomponents was seen to correspond to the
pattern found in adult livers when reacted with both antisera
(Pattern G, Chart 5).
Assay of Hepatic Catatase
Catalase activity was assayed in each of the aforementioned
liver homogenates from fetal, newborn, and adult rats. The
levels of catalase activity expressed in mg nitrogen of the samples
or ¿To/Nare graphically shown in Chart 6. The graph illustrates
a gradual increase in enzyme activity which reaches a plateau
in the perinatal period and recedes slightly in the 14-day newborn
period before reaching the adult levels at 25 days.
Morphologic Studies of Fetal and Xewborn Rat Liver
PURE
REC
CHART 4. Immunoelectrophoretic patterns of purified rat
erythrocyte catalase reacted against rabbit anti-rat erythrocyte
catalase serum (AbREC) and rabbit anti-rat liver catalase serum.
Compare with Chart 1.
Morphologic studies of organogénesisby light microscopy
obviously provide little direct evidence of intracellular modifica
tions during maturation and growth. These studies, do, however,
provide means for the evaluation of cell types and allow for the
assessment of degrees of structural variations present during
different developmental periods of the organ under study. It,
therefore, seemed pertinent to include a morphologic analysis as
NEWBORN
FETAL
NEWBORNNEWBORN ADULT
24 hr
14 d
(&25DAY
N.B.)
FETUS
16 d
A
ADULT
B
C
D
E
F
G
CHART5. Immunoelectrophoretic patterns for various stages of development. Patterns are contrasted between reactions of rabbit
anti-rat liver catalase and rabbit anti-rat erythrocyte catalase sera. Diagrams of imnumoelectrophoretic patterns of fetal, newborn,
and adult liver homogenates reacted with AbRHC and AbREC.
120
CANCER RESEARCH VOL. 27
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Changes of Hepatic Calalase
1.6
HEPATIC
CATALASE
ACTIVITY
IN FETAL AND NEWBORN RATS
3S 1.4
¿1.2
LO
0.8
0.6
N
0.4
0.2
I
16
I
18 20
FETUS
I
I
I
I
I
22 : I
5
10
\5
20
BIRTH
NEWBORN
AGE IN DAYS
J_
25
JL
160
ADULT
CHAUT6. Changes in enzyme activity of liver homogenates during embryogenesis. Note the plateau between birth and 15th postpart urn day.
SUMMARY OF HEPATIC CATALASEDEVELOPMENT
n(t
STAGEIMMUNOELEC-
(fov
Vr
TROPHORESIS
RHC)AGE:ACTIVITYKo/N
(Ab
(16df
NB -
19-21df.146
ADULT1.20
NB.669
.407
%
ADULTLIVER
100%OERYTHROPOIESISADULT
36%+++ERYTHROPOIESIS0
12%
56%++ERYTHROPOIESISEARLY
50%¿ERYTHROPOIESISADULT
MORPHOLOGY:i
STRUCTURE0
CORD
PORTAL
TRIADSi
CORDFORMA
TIONEARLY
CORD STRUC
CORD STRUC
TUREADULT
TUREADULT
PORTAL
PORTAL
TRIADSmTJ('•14dNB.594
TRIADSWY(f°(Õ25d
PORTAL
TRIADSY
nmt22df-22hr
CHART7. Summary chart correlating the imrmmoeleetrophoretic patterns with catalase activity and morphologic development
of the liver.
a 3rd parameter in order to attempt a correlation between
morphology and ontogenesis of catalase isozymes.
Liver development during the ages studied in this experiment
falls into 4 categories. The 1st stage includes fetal rats with
ages of 16-21 days. Little structural organization of the liver is
apparent at this stage, and marked erythropoiesis is noted. The
22-day fetus and early-newborn livers demonstrate early and
JANUARY
irregular hepatic cord formation and reduced erythropoiesis. The
14-day newborn liver reveals structural development which
closely approaches the adult form. Hepatic lobules with orderly
cord formation and portal triads are present, and only occasional
clumps of immature erythrocytes are observed. The 4th stage
consists of typical adult rat liver pattern with no evidence of
hematopoiesis.
1967
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121
Grant W. Pation and E divin T. Nishimiira
Discussion
A summary of the qualitative and quantitative analyses of
hepatic catalase and comments on the corresponding liver
morphology during the fetal and newborn period of development
are presented in Chart 7. Qualitative changes in the enzyme
established by immunoelectrophoresis revealed at least 5 stages
which precede the ultimate formation of adult-type hepatic
catalase during embryonic and perinatal development. Interest
ing aspects of the enzyme modification included the shift from a
predominantly anodic to the cathodic arcs and allowed, to some
extent, a correlation of these subcomponents with changes in
enzyme activity. The appearance of the long cathodic arc in
Stage II was associated with a 3-fold increase in enzyme activity;
however, a unique anodic arc was also observed at this time.
Stage III demonstrated little change in the subcomponent
pattern to account for the 20% increase in enzyme activity. Of
perhaps greater significance was the marked change in pattern
during Stage IV. Hepatic catalase activities assayed on speci
mens from 5 littermates of the 14-day newborn period revealed a
wide variation in A^/N values ranging from 0.718 to 0.311 as
recorded in Chart 6. Immunoelectrophoretic pattern of all
samples of the littermates revealed arc configuration presented
in Chart 3. The mean value of these activities was below the
early newborn level and correlated well with data of the newborn
catalase levels presented by Stevens (18). Evidence reported
earlier from starvation and tumor studies (10) suggests that total
body iron may be deficient during the newborn period and is thus
shunted into other metabolic pathways, resulting in decreased
production of catalase. Although the data to support the concept
are lacking, it is tempting to speculate on the possibility that
some factor(s) involved with the changing developmental or
environmental conditions may have "turned off" the regulating
factor responsible for synthesis of Subcomponents H-4 and H-5
(repression?). It is interesting too that the nestling period for rats
ends at about 15-16 days, and perhaps this, in part, may be the
cause or the result of the enzymatic changes observed.
Extramedullary erythropoiesis in fetal and newborn rat livers
has been previously described and was observed in this study.
One would expect quantitative changes in erythrocyte catalase
which undoubtedly coexists with hepatic catalase during the
maturation process. The estimate of the amount of erythrocyte
catalase in the liver as demonstrated by immunoelectrophoresis
of liver homogenates with AbREC failed to reveal any definite
alterations in the subcomponent pattern of erythrocyte catalase.
This result was not unexpected in view of the low concentration
of erythrocyte catalase per mass of RBC protein and the rela
tively low proportion of RBC to liver cells in these samples.
Moreover, immature RBC may possess low catalatic activity and
contribute little in this regard.
The modifications of pattern in liver catalase during embryogenesis suggest that fetal catalase may undergo serial differentia
tion which corresponds to specific functional needs of the liver
tissues during maturation. Deficit of arcs observed in the very
young fetal tissues may reflect the immature state of cell develop
ment and, therefore, the inability of such cells to synthesize all
the necessary subcomponents of the catalase molecule. A study
of the ultrastructure of differentiating hepatic cells which corre
122
spond to various stages of catalase-subcomponent development
may shed some additional light on the problem.
The heterogeneous quality of catalase and the complex modi
fications it undergoes during early fetal and newborn life may
have some relationship to the rate of cell proliferation. Quantita
tive changes of catalatic activity of neoplastic cells may be
related to a modification in the molecular makeup of catalase.
Repression of liver catalase synthesis during extra-hepatic tumor
growth, likewise, may be a reflection of disordered metabolic
state. Such changes in catalase activity during neoplastic growth
may be related to dedifferentiation of cells (or cell ultrastructure)
and, hence, may indicate reversion of catalase subcomponents to
a more primitive or fetal pattern. Investigation is currently
underway to compare the patterns of liver catalase in non-tumorbearing and tumor-bearing hosts.
Acknowledgments
The authors are indebted to Mr. Stanley N. Carson for his cap
able assistance on this project.
References
1. Agner, K. The Preparation and Properties of a Highly Active
Catalase from Horse Liver. Biochem. J., 32: 1702-6, 1938.
2. Beckman, L., Scandalios, J. G., andBrewbaker, J. L. Catalase
Hybrid Enzymes in Maize. Science, 146:1174-75, 1964.
3. Bonnichsen, R. K. A Comparative Study of the Blood and
Liver Catalases from the Horse. Arch. Biochem. Biophys.,
le: 83-94, 1947.
4. Brown, G. L. A Solubility Analysis of Crystalline Ox-Liver
Catalase. Biochem. J., 51: 569-73, 1952.
5. Clayton, R. K. High-Catalase Mutant Rhodopseudomonan
Spheroides.
Characterization
of the Catalase.
Biochim.
Biophys. Acta, 58: 360-62, 1962.
6. Deutsch, H. F., and Scabra, A. Immunochemical Studies and
Assay of Catalase. J. Biol. Chem., 214: 455-62, 1955.
7. Greenstein, J. P. The Liver Catalase Activity of Tumorbearing Mice and the Effect of Spontaneous Regression and of
Removal of Certain Tumors. J. Nati. Cancer Inst., %:345-55,
1942.
8. Higashi, T., and Peters, T. Studies on Rat Liver Catalase. I.
Combined Immunochemical and Enzymatic Determination of
Catalase in Liver Cell Fractions. J. Biol. Chem., 338: 3945-51,
1963.
9. Higashi, T., Yagi, M., and Hirai, H. Immunochemical Studies
on Catalase. J. Biochem. Tokyo, 49: 707-12, 1961.
10. Nakahara, W., and Fukuoka, F. Chemistry of Cancer Toxin:
Toxohormone.
Springfield,
111.: Charles C Thomas Pub
lisher, 1961.
11. Nakatani, M. A Chromatographie Study of Crystalline Cata
lase from Bovine Liver. Bull. Agr. Chem. Soc. Japan, 23:
308-13, 1958.
12. Nishimura, E. T., Carson, S. N., and Kobara, T. Y. Isozymes
of Human and Rat Catalases. Arch. Biochem. Biophys., 108:
452-59, 1964.
13. Nishimura, E. T., Carson, S. N., and Patton, G. W. Immuno
electrophoretic
Identification of Catalase Subcomponents in
the Homogenates of Rat Tissues. Cancer Res., 26: 92-96, 1966.
14. Saha, A., Campbell, D. H., and Schroeder, W. A. Immuno
chemical Studies on Liver and Erythrocyte
Catalases from
Cattle, Horse, Rabbit and Human. Biochim. Biophys. Acta,
85: 38-49, 1964.
CANCER RESEARCH VOL. 27
Downloaded from cancerres.aacrjournals.org on June 15, 2017. © 1967 American Association for Cancer Research.
Changes of Hepatic Catalase
15. Samejima, T. Splitting of Catalase Molecule by Alkali Treatment. J. Biochem. Tokyo, 46: 155-59, 1959.
16. Samejima, T., and Shibata, K. Denaturation
of Catalase by
Formamide
and
.
,
, Urea
. , Related to, the Subunit Make-upr of the
Molecule. Arch. Biochem. Biophys., 93: 407-12, 1961.
17. Schroeder, W. A., Shelton, J. R., Shelton, J. B., and Olson, B.
M. Some Amino Acid Sequences in Bovine-Liver Catalase.
Biochim. Biophys. Acta, 89; 47-65, 1964.
18. Stevens, L. A Comparative
Study of Enzymes in Foetal,
JANUARY
Young and Adult Rat. Comp. Biochem. Physiol., 6: 129-35,
1962.
19. Sumner, J. B., and Grälen, N. The Molecular Weight of
„
. „. r>
* i * J.T -nion QQQ«
Crystalline
Catalase.
Biol.i r«i*
Chem., IzB:
33-36, ima
1938.
W- Tanford, C., and Lovrien, R. Dissociation of Catalase into
Subunits. J. Am. Chem. Soc., 84: 1892-96, 1962.
21. von Euler, H., and Josephson, K. Ãœber Katalase I. Ann.
Chem., 4BS: 158-81, 1927.
1967
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123
Developmental Changes of Hepatic Catalase in the Rat
Grant W. Patton and Edwin T. Nishimura
Cancer Res 1967;27:117-123.
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