- Email: [email protected]
Abolition of the crabtree effect in Ehrlich ascites tumor cells by vitamin K3
aud ecluilibraticm, iujectiou, and also in uilrv.
shows coutains
the highest tnitocliondria
._c a, 0 h
50
3L
1 10
iudebted
cone
to
0T acetaltleti~dc seusitivity
after :dcohol to acrtaldeliyde
5.0
Acetaldehyde
I am
highest
I.‘io h’ 1. -Effect of acetaltkh~tk on ositlation of pyruvate \vith mitnchontlria from different parts of rat brain. Hrspiration was measuretl by the conventional \Varburg method with air as the gas phase ant1 with 0.3 ml of 2 .\I potassium h~drositle in the renter H~cII. The illcubations were performed at 30 (: and lastetl for 20 min. Each vessel containetl 1 ml of mitochondria (corresponding to 301) mg tresh tissue), 311 pmoles pyruvate, 63 . ., ,umolrs potassium phosphate (pH 7.5). 4.5 ;tmoles A’TI’. 15 ymoles magnesium sulphate, 4.5 /&moles glucose, 1 mg he\-okinaw (Sigma, gratle III), and 150 pmoles sucrose (besides that already adclett with the mitochontlrla). Xcetaldehyde. when indicated, was atlclrtl just before the rsperimcnt was started. Final volume 2.3 ml.
-
F L
couccntration with the
mM
Miss
PvIargareta
work is part of investigations Science Research Council.
made
Skoglund possible
for skilful technical assistance. This by grants from the Swedish Medical
REFERENCES 1. 2. 3. 4.
BEER, C. T. and QUASTEL, .I. H., (:ctn. .I. Riochem. and Physiol. BRODY, T. RI. and BAIN, J. A., .I. Riol. Chem. 195, 685 (1952). KIESSLING, K.-H., Esptl. Cell Research 26, 432 (1962). STOTZ, E., J. Riol. Chem. 148, 585 (1913).
ABOLITION
OF THE
CRABTREE
TUMOR
EFFECT
IN EHRLICH
CELLS BY VITAMIN
G. DALLNER
531, 36 (1958).
K,
and I,. ERNSTER
Kenner-Gren Institute, University of Stockholm, and Institute Sabbatsberg Hospital, Karolinska Institutet, Stockholm, Received
MALIGNANT
ASCITES
May
of Pathology
at
Sweden
7, 1962
tumors, as well as certain normal tissues with high aerobic glycolysis, exhibit diminished respiration when fermentable monosaccharides such as glucose, fructose and mannose are added. This form of respiratory inhibition is known as the Crabtree [6] or reversed Pasteur effect. The inhibition can be abolished by the addition of uncouplers of oxidative phosphorylation, such as 2&dinitrophenol [12]. This Experimental
Cell
Reseurch
27
Abolition
of the Crabtree effect by oitamin
K,
suggests the dependence of the Crabtree effect on respiratory control, that is, on a respiration which is tightly coupled to phosphorylation. According to the generally accepted view (for review, see [l]), the reason for the Crabtree effect is a competition for inorganic phosphate and/or phosphate acceptor between a part of the glycolytic svstem and the energy generating mitochondrial respiration. 2-Deoxyglucose cannot b
-log
M vitamin
Kg
Dicoumarol.
M = 106
Pig. I.--Effect of vitamin K, (n) and dicoumarol (b) on the respiration of Ehrlich ascites tumor cells. Each vessel contained 1.2 ml Krehs-Ringer-phosphate solution, pH 7.4, without Ca”+, and 0.4 ml ascites tumor cell suspension (11.0 mg protein in esp. o and 15.8 mg protein in exp. b) in a final volume of 2 ml. \Vhen indicated, 0.01 JI glucose, varying concentrations (a) or 3 x 10-S J\I (b) vitamin K,, and varying concentrations of dicoumarol were added. Incubation at 37°C with air as gas phase.
undergo glycolysis, although it is capable of being phosphorylated through the hexokinase reaction, and yet it elicits a respiratory inhibition [ill. This finding would apparently eliminate the intact glycolytic system from participating in the Crabtree effect, and thus would localize the effect to the hexokinase reaction. Direct experimental proof, however, for the above hypotheses is not yet available. A promising new approach to this problem has now become possible by the use of vitamin I<, (2-methyl-l,&naphthoquinone) as an experimental tool. It has been shown earlier [5, 91 that in rat liver mitochondria added vitamin I<, can establish an electron shunt between DPNH’ and cytochrome 6, i.e., over that portion of the respiratory chain which is inhibited by Amytal, and where the first of the three phosphorylations occurs; this part of the electron transport system is generally accepted as being rate-limiting during phosphate- and/or phosphate-acceptor-controlled respiration. The above “vitamin I(, shunt” was shown to involve the pyridine nucleotide nonspecific flavoenzyme, DT diaphorase [lo], which is characterized by a high sensitivity to dicoumarol. Since it had been found [S] that ascites tumor cells contain DT diaphorase, an investigation of the effect of vitamin I<, on the respira1 -4bbreviations: nucleotide.
DPN,
diphosphopyridine
nucleotide;
DPNH,
reduced
Experimental
diphosphopyridine
Cell Reseurch
27
370 tion of these cells appeared to be of interest in connection \vith our studies OCthe mechanism of the Crabtrec effect. In all esperiments E&lich (ELT) asrites tumor cells were pooled from hct~~rozygous albino mice 11 days after inoculation. The cells were washed 2 3 times by low speed centrifugation in an International Kefrigerated Centrifuge to remove erythrocytes, before packing them by ceutrifugation at 1500 rpm for 5 min. The cell mass was then suspended in two volumes of medium. The washing and suspending medium consisted of Iirebs-Ringer-phosphate solution, pH 7.4, without Ca”+. Respiration was measured by the \Varburg method. Vitamin I<, in concentrations exceeding 1W6 JI exhibited a pronounced effect on the respiration of the Ehrlich ascites tumor cells (Fig. 1 n). In the presence of glucose, the Crabtree effect was abolished. The same vitamin Ii, concentrations caused a depression of the endogenous respiration. The vitamin Ii, activation of the respiration in the presence of glucose could be largely removed by 1O-6 A1 dicoumarol (Fig. 1 h), indicating the involvement of DT diaphorase. Higher concentrations of dicoumarol stimulated the oxygen consumption both without and with @LLCcJSe, and also with glucose plus vitamin I<,, but did not change the endogenous respiration depressed by vitamin K,. This dicoumarol stimulation is clearly a consequence of the dissociation of the respiration from phosphorylation, inasmuch as a similar effect ensued with dinitrophenol. Other naphthoquinones, as well as various benzoquinones, did not share the effect of vitamin I<, in abolishing the Crabtree effect (Table I). This finding is in accordance with the previously [5, 9] established specificity of the “vitamin K, shunt” for vitamin Ii,. Replacement of glucose by fructose or mannose did not alter the vitamin I<, effect, whereas with 2-deoxyglucose, the ensuing depression of the respiration was only partially restored by vitamin I<, (Table II). Amytal (2 m&l), which inhibited the respiration in the absenceof vitamin I(, by X0-90 per cent both in the absenceand presenceof added hexose, gave in the presenceof vitamin I<, only about 50 per cent inhibition of the respiration with endogenoussubstrate (with or without 2-deosgglucose),and only about 20 per cent with glucose,fructose or mannose. It thus appears that the “vitamin I(, shunt” is more efficient with substrates, such as the fermentable hexoses, which can give rise to extramitochondrial DPNH, than it is with the endogenous substrates-probably fatty acids [13]---whose oxidation proceeds mainly or exclusively by way of intramitochondrial DPN. This finding is probably related to the fact, established in preliminary experiments by direct assay of the isolated cell fractions, that the bulk of the DT diaphorase in the tumor cells is located outside the mitochondria, and can thus only operate with extramitochondrial pyridine nucleotides as electron donors. X similar situation has earlier been found in rat liver 14,91. In the glucose-supplementedsystem, the medium levels of glucoseand lactate were also followed (see footnote to Table II). The accumulation of lactate increased in the presence of Xmytal. Under all conditions, the sum of the oxygen consumption and lactate accumulation was roughly equal to twice the amount of glucoseconsumed, indicating that practically all respiration proceeded by oxidation of glycolytically generated DPNEI. The finding that, in spite of this, the respiration was almost completely inhibited by Amytal suggeststhat the aerobic oxidation of extramitochondrial DPNH in the ascites tumor cells proceeds mainly or exclusively by way Erperimenfal
Cell Reseurch
27
Abolition TABLE
I.
Effect of various
of the Crabtree effect by vitamin quinones
on the respiration
371
K,
of Ehrlich
ascites tumor
cells.
Conditions as in Fig. 1. 0.4 ml ascites tumor cell suspension, containing 20.3 mg protein per vessel. Quinones were added in a final concentration of 3 x 1O-5 -11, with the exception of coenzyme Qz, which was 10-S -11. Incubation at 37’C for 30 min. With Added
glucose patorns
quinone
None Vitamirl K, l,l-naphthoquinone Z-methyl-3-OH-naphthoquinone 2-OH-1.4naphthoquinone Vitamin Ii, Vitamin Ii, Synkavit Vitamin E 1 A-benzoquinone 2-methyl-l,l-benzoquinone 2,6-dimethyl-1.4benzoquinone Coenzyme Q,, Coenzyme Qa Coenzyme Q6 Coenzyme Qs Coenzyme Qlo
TABLE
Without oxygen
glucose
6.8
7.7 5.8
3.3
3.0
4.5
7.5
4.1
i.3
3.6
7.5
4.0
7.5
4.2
7.7
3.5
6.8
3.6
i.3
3.6
i.3
3.5 3.5
7.0 -
-4.2
6.5
3.5
i.2
3.6
3.8
5.4
3.5
6.8
II. Effect of vitamin K3 and Amytal on the respiration of Ehrlich cells in the presence of various hexoses.
Conditions as in Fig. 1. Hesoses were added in concentrations of 0.01 cell suspension, containing 20.1 mg protein per vessel. When indicated, 3 >. 10-h Jf vitamin K, were added. Incubation at 37°C for 30 min. Without without Added
vit.
hesose
K,
Amytal with
JI. 0.4 ml ascites tumor 0.002 .1I Amytal and
With vit. Ii, ,Uatoms
without osygen
ascites tumor
rit.
;Imytal I<,
with
vit.
None
8.8
5.8
0.5
2.2
Glucosea
1.9
8.6
0.9
6.0
Rfannose Fructose
1.3
7.0
0.9
5.5
5.2
8.4
0.9
5.8
2-deoxyglucose
3.5
4.8
0.6
2.8
K,
a The four consecutive samples indicated in this line, when analyzed after incubation, had consumed 10.0, 10.1, 11.9 and 11.0 pmoles glucose, and had accumulated 13.6, 12.8, 20.0 and 14.4 pmoles L-lactate. E.rperimenlal
Cell
Research
27
of intralnitocholldrial DPK;, probably involving a substrate-c~ltal?zetl, cyclic transfcl of hydrogen [:
Crabtree effect, obtained in esperiments will
be reported
in a forthcoming
taking advantage of the present approach,
communication
171.
This work has been supported by a grant from the Swedish Cancer Society. The valuable assistance of Mrs. Kerstin Krantz is gratefully acknowledged. REFERENCES 1. XISENBERG, -1. C., The Glycolysis ant1 Respiration of Tumors. 1961. 2. BORST, P., Biochirn. et Biophys. dcfo 57, 250 (196’L). 3. BOXER, G. E. and DE~LIS, T. RI., Science 134, 1495 (1961).
1. CONOVER,
T. E.,
in Biolo@cal
Structure
and Funrtion,
vol.
Acadetnic
Press,
Sew
2, p. 169. Gooowrx,
York,
T. \V. and
LINDBERG, 0. (eds.j.Xcademic Press, L.ondon, 1961. 5. CONOVER, T. E. and ERNSTER, L., Biochim. et Biophgs. dcln 58, 189 (1962). 6. CRABTREE, H. G., Rio&em. J. 23, 536 (1929). 7. DALLNER, G. and ERNSTER, L., fhpII. Cell Krsearch 27. 352 (1962). 8. ERNSTER, L., Federution Proc. 17, 216 (1958). 9. ERWTER, I.., in Biological Structure ant1 Functiou, vol. 2. p. 139. Goar~wr~, T. IV. aud LINUBERG, 0. (eds.). Academic Press. London. 1961. 10. ERNSTER, L., DANIELSON, L. and LJUP~GGREN, Ri., Biochim. el Biophys. dclu 58, 171 (1962). 11. IBSEN, I<. H., (;OE, E. L. aIltl ikf&EE, B. \I’., BiOChiIIL et BiOphIJS. dCkI 30, 381 (19%). 1’. Itus, E., T.~L.~LAY, P. autl \VILLI.~MS-.\SH~U~N. H. G., Cancer Research 11, 855 (1951). 13. MEDES, G. and WEINHOUSE, S. Cn~Icw Kesenrch 18, 352 (195X).
INDUCTION
OF A CRABTREE-LIKE ASCITES
TUMOR G. DALLNER
EFFECT
CELLS and
BY
IN
a preceding
communication
May
[-4] we have
EHRLICH
OLIGOMYCIN
L. ERNSTER
W’enner-Gren Institute, UniLwsity of Stockholm, and Institute Sabbatsberg Hospital, Karolinska Institutet, Stockholm, Receiwd
IN
of Pathology Sweden
at
11: 1962 presented
evidence
that
the
depression
of
tumor cell respiration by hexoses, known as the Crabtree effect [3], is reflected in a rate limitation of mitochondrial electron transfer between DPN’ and cytochrome b. 1 Abbreviations: nucleotide.
Experimental
DPN,
Cell Research
diphosphopgridine
27
nucleotide;
TPNH,
reduced
triphosphopyridine
shows coutains
the highest tnitocliondria
._c a, 0 h
50
3L
1 10
iudebted
cone
to
0T acetaltleti~dc seusitivity
after :dcohol to acrtaldeliyde
5.0
Acetaldehyde
I am
highest
I.‘io h’ 1. -Effect of acetaltkh~tk on ositlation of pyruvate \vith mitnchontlria from different parts of rat brain. Hrspiration was measuretl by the conventional \Varburg method with air as the gas phase ant1 with 0.3 ml of 2 .\I potassium h~drositle in the renter H~cII. The illcubations were performed at 30 (: and lastetl for 20 min. Each vessel containetl 1 ml of mitochondria (corresponding to 301) mg tresh tissue), 311 pmoles pyruvate, 63 . ., ,umolrs potassium phosphate (pH 7.5). 4.5 ;tmoles A’TI’. 15 ymoles magnesium sulphate, 4.5 /&moles glucose, 1 mg he\-okinaw (Sigma, gratle III), and 150 pmoles sucrose (besides that already adclett with the mitochontlrla). Xcetaldehyde. when indicated, was atlclrtl just before the rsperimcnt was started. Final volume 2.3 ml.
-
F L
couccntration with the
mM
Miss
PvIargareta
work is part of investigations Science Research Council.
made
Skoglund possible
for skilful technical assistance. This by grants from the Swedish Medical
REFERENCES 1. 2. 3. 4.
BEER, C. T. and QUASTEL, .I. H., (:ctn. .I. Riochem. and Physiol. BRODY, T. RI. and BAIN, J. A., .I. Riol. Chem. 195, 685 (1952). KIESSLING, K.-H., Esptl. Cell Research 26, 432 (1962). STOTZ, E., J. Riol. Chem. 148, 585 (1913).
ABOLITION
OF THE
CRABTREE
TUMOR
EFFECT
IN EHRLICH
CELLS BY VITAMIN
G. DALLNER
531, 36 (1958).
K,
and I,. ERNSTER
Kenner-Gren Institute, University of Stockholm, and Institute Sabbatsberg Hospital, Karolinska Institutet, Stockholm, Received
MALIGNANT
ASCITES
May
of Pathology
at
Sweden
7, 1962
tumors, as well as certain normal tissues with high aerobic glycolysis, exhibit diminished respiration when fermentable monosaccharides such as glucose, fructose and mannose are added. This form of respiratory inhibition is known as the Crabtree [6] or reversed Pasteur effect. The inhibition can be abolished by the addition of uncouplers of oxidative phosphorylation, such as 2&dinitrophenol [12]. This Experimental
Cell
Reseurch
27
Abolition
of the Crabtree effect by oitamin
K,
suggests the dependence of the Crabtree effect on respiratory control, that is, on a respiration which is tightly coupled to phosphorylation. According to the generally accepted view (for review, see [l]), the reason for the Crabtree effect is a competition for inorganic phosphate and/or phosphate acceptor between a part of the glycolytic svstem and the energy generating mitochondrial respiration. 2-Deoxyglucose cannot b
-log
M vitamin
Kg
Dicoumarol.
M = 106
Pig. I.--Effect of vitamin K, (n) and dicoumarol (b) on the respiration of Ehrlich ascites tumor cells. Each vessel contained 1.2 ml Krehs-Ringer-phosphate solution, pH 7.4, without Ca”+, and 0.4 ml ascites tumor cell suspension (11.0 mg protein in esp. o and 15.8 mg protein in exp. b) in a final volume of 2 ml. \Vhen indicated, 0.01 JI glucose, varying concentrations (a) or 3 x 10-S J\I (b) vitamin K,, and varying concentrations of dicoumarol were added. Incubation at 37°C with air as gas phase.
undergo glycolysis, although it is capable of being phosphorylated through the hexokinase reaction, and yet it elicits a respiratory inhibition [ill. This finding would apparently eliminate the intact glycolytic system from participating in the Crabtree effect, and thus would localize the effect to the hexokinase reaction. Direct experimental proof, however, for the above hypotheses is not yet available. A promising new approach to this problem has now become possible by the use of vitamin I<, (2-methyl-l,&naphthoquinone) as an experimental tool. It has been shown earlier [5, 91 that in rat liver mitochondria added vitamin I<, can establish an electron shunt between DPNH’ and cytochrome 6, i.e., over that portion of the respiratory chain which is inhibited by Amytal, and where the first of the three phosphorylations occurs; this part of the electron transport system is generally accepted as being rate-limiting during phosphate- and/or phosphate-acceptor-controlled respiration. The above “vitamin I(, shunt” was shown to involve the pyridine nucleotide nonspecific flavoenzyme, DT diaphorase [lo], which is characterized by a high sensitivity to dicoumarol. Since it had been found [S] that ascites tumor cells contain DT diaphorase, an investigation of the effect of vitamin I<, on the respira1 -4bbreviations: nucleotide.
DPN,
diphosphopyridine
nucleotide;
DPNH,
reduced
Experimental
diphosphopyridine
Cell Reseurch
27
370 tion of these cells appeared to be of interest in connection \vith our studies OCthe mechanism of the Crabtrec effect. In all esperiments E&lich (ELT) asrites tumor cells were pooled from hct~~rozygous albino mice 11 days after inoculation. The cells were washed 2 3 times by low speed centrifugation in an International Kefrigerated Centrifuge to remove erythrocytes, before packing them by ceutrifugation at 1500 rpm for 5 min. The cell mass was then suspended in two volumes of medium. The washing and suspending medium consisted of Iirebs-Ringer-phosphate solution, pH 7.4, without Ca”+. Respiration was measured by the \Varburg method. Vitamin I<, in concentrations exceeding 1W6 JI exhibited a pronounced effect on the respiration of the Ehrlich ascites tumor cells (Fig. 1 n). In the presence of glucose, the Crabtree effect was abolished. The same vitamin Ii, concentrations caused a depression of the endogenous respiration. The vitamin Ii, activation of the respiration in the presence of glucose could be largely removed by 1O-6 A1 dicoumarol (Fig. 1 h), indicating the involvement of DT diaphorase. Higher concentrations of dicoumarol stimulated the oxygen consumption both without and with @LLCcJSe, and also with glucose plus vitamin I<,, but did not change the endogenous respiration depressed by vitamin K,. This dicoumarol stimulation is clearly a consequence of the dissociation of the respiration from phosphorylation, inasmuch as a similar effect ensued with dinitrophenol. Other naphthoquinones, as well as various benzoquinones, did not share the effect of vitamin I<, in abolishing the Crabtree effect (Table I). This finding is in accordance with the previously [5, 9] established specificity of the “vitamin K, shunt” for vitamin Ii,. Replacement of glucose by fructose or mannose did not alter the vitamin I<, effect, whereas with 2-deoxyglucose, the ensuing depression of the respiration was only partially restored by vitamin I<, (Table II). Amytal (2 m&l), which inhibited the respiration in the absenceof vitamin I(, by X0-90 per cent both in the absenceand presenceof added hexose, gave in the presenceof vitamin I<, only about 50 per cent inhibition of the respiration with endogenoussubstrate (with or without 2-deosgglucose),and only about 20 per cent with glucose,fructose or mannose. It thus appears that the “vitamin I(, shunt” is more efficient with substrates, such as the fermentable hexoses, which can give rise to extramitochondrial DPNH, than it is with the endogenous substrates-probably fatty acids [13]---whose oxidation proceeds mainly or exclusively by way of intramitochondrial DPN. This finding is probably related to the fact, established in preliminary experiments by direct assay of the isolated cell fractions, that the bulk of the DT diaphorase in the tumor cells is located outside the mitochondria, and can thus only operate with extramitochondrial pyridine nucleotides as electron donors. X similar situation has earlier been found in rat liver 14,91. In the glucose-supplementedsystem, the medium levels of glucoseand lactate were also followed (see footnote to Table II). The accumulation of lactate increased in the presence of Xmytal. Under all conditions, the sum of the oxygen consumption and lactate accumulation was roughly equal to twice the amount of glucoseconsumed, indicating that practically all respiration proceeded by oxidation of glycolytically generated DPNEI. The finding that, in spite of this, the respiration was almost completely inhibited by Amytal suggeststhat the aerobic oxidation of extramitochondrial DPNH in the ascites tumor cells proceeds mainly or exclusively by way Erperimenfal
Cell Reseurch
27
Abolition TABLE
I.
Effect of various
of the Crabtree effect by vitamin quinones
on the respiration
371
K,
of Ehrlich
ascites tumor
cells.
Conditions as in Fig. 1. 0.4 ml ascites tumor cell suspension, containing 20.3 mg protein per vessel. Quinones were added in a final concentration of 3 x 1O-5 -11, with the exception of coenzyme Qz, which was 10-S -11. Incubation at 37’C for 30 min. With Added
glucose patorns
quinone
None Vitamirl K, l,l-naphthoquinone Z-methyl-3-OH-naphthoquinone 2-OH-1.4naphthoquinone Vitamin Ii, Vitamin Ii, Synkavit Vitamin E 1 A-benzoquinone 2-methyl-l,l-benzoquinone 2,6-dimethyl-1.4benzoquinone Coenzyme Q,, Coenzyme Qa Coenzyme Q6 Coenzyme Qs Coenzyme Qlo
TABLE
Without oxygen
glucose
6.8
7.7 5.8
3.3
3.0
4.5
7.5
4.1
i.3
3.6
7.5
4.0
7.5
4.2
7.7
3.5
6.8
3.6
i.3
3.6
i.3
3.5 3.5
7.0 -
-4.2
6.5
3.5
i.2
3.6
3.8
5.4
3.5
6.8
II. Effect of vitamin K3 and Amytal on the respiration of Ehrlich cells in the presence of various hexoses.
Conditions as in Fig. 1. Hesoses were added in concentrations of 0.01 cell suspension, containing 20.1 mg protein per vessel. When indicated, 3 >. 10-h Jf vitamin K, were added. Incubation at 37°C for 30 min. Without without Added
vit.
hesose
K,
Amytal with
JI. 0.4 ml ascites tumor 0.002 .1I Amytal and
With vit. Ii, ,Uatoms
without osygen
ascites tumor
rit.
;Imytal I<,
with
vit.
None
8.8
5.8
0.5
2.2
Glucosea
1.9
8.6
0.9
6.0
Rfannose Fructose
1.3
7.0
0.9
5.5
5.2
8.4
0.9
5.8
2-deoxyglucose
3.5
4.8
0.6
2.8
K,
a The four consecutive samples indicated in this line, when analyzed after incubation, had consumed 10.0, 10.1, 11.9 and 11.0 pmoles glucose, and had accumulated 13.6, 12.8, 20.0 and 14.4 pmoles L-lactate. E.rperimenlal
Cell
Research
27
of intralnitocholldrial DPK;, probably involving a substrate-c~ltal?zetl, cyclic transfcl of hydrogen [:
Crabtree effect, obtained in esperiments will
be reported
in a forthcoming
taking advantage of the present approach,
communication
171.
This work has been supported by a grant from the Swedish Cancer Society. The valuable assistance of Mrs. Kerstin Krantz is gratefully acknowledged. REFERENCES 1. XISENBERG, -1. C., The Glycolysis ant1 Respiration of Tumors. 1961. 2. BORST, P., Biochirn. et Biophys. dcfo 57, 250 (196’L). 3. BOXER, G. E. and DE~LIS, T. RI., Science 134, 1495 (1961).
1. CONOVER,
T. E.,
in Biolo@cal
Structure
and Funrtion,
vol.
Acadetnic
Press,
Sew
2, p. 169. Gooowrx,
York,
T. \V. and
LINDBERG, 0. (eds.j.Xcademic Press, L.ondon, 1961. 5. CONOVER, T. E. and ERNSTER, L., Biochim. et Biophgs. dcln 58, 189 (1962). 6. CRABTREE, H. G., Rio&em. J. 23, 536 (1929). 7. DALLNER, G. and ERNSTER, L., fhpII. Cell Krsearch 27. 352 (1962). 8. ERNSTER, L., Federution Proc. 17, 216 (1958). 9. ERWTER, I.., in Biological Structure ant1 Functiou, vol. 2. p. 139. Goar~wr~, T. IV. aud LINUBERG, 0. (eds.). Academic Press. London. 1961. 10. ERNSTER, L., DANIELSON, L. and LJUP~GGREN, Ri., Biochim. el Biophys. dclu 58, 171 (1962). 11. IBSEN, I<. H., (;OE, E. L. aIltl ikf&EE, B. \I’., BiOChiIIL et BiOphIJS. dCkI 30, 381 (19%). 1’. Itus, E., T.~L.~LAY, P. autl \VILLI.~MS-.\SH~U~N. H. G., Cancer Research 11, 855 (1951). 13. MEDES, G. and WEINHOUSE, S. Cn~Icw Kesenrch 18, 352 (195X).
INDUCTION
OF A CRABTREE-LIKE ASCITES
TUMOR G. DALLNER
EFFECT
CELLS and
BY
IN
a preceding
communication
May
[-4] we have
EHRLICH
OLIGOMYCIN
L. ERNSTER
W’enner-Gren Institute, UniLwsity of Stockholm, and Institute Sabbatsberg Hospital, Karolinska Institutet, Stockholm, Receiwd
IN
of Pathology Sweden
at
11: 1962 presented
evidence
that
the
depression
of
tumor cell respiration by hexoses, known as the Crabtree effect [3], is reflected in a rate limitation of mitochondrial electron transfer between DPN’ and cytochrome b. 1 Abbreviations: nucleotide.
Experimental
DPN,
Cell Research
diphosphopgridine
27
nucleotide;
TPNH,
reduced
triphosphopyridine