An acceptor effect in glycolysis

Vol. 1, No. 3

BfOCHEMfCAl

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

AN ACCEPTOR

EFFECT

Jan van Eys*

August

In order especially order

in liver,

to inhibit

using

the loss

used

in oxidative acceptor

system.

lation,

2 -deoxyglucose

mouse

liver

This

technique

of an acceptor

system,

couple

might

rather

-6 -phosphate

does

the product

, is a relatively

poor

cannot

that it inhibits

of residual

via

in a

substrate

for

advantage

that

further,

but

be metabolized

glycogen

result

be

of phosphory-

It has the additional

-6-phosphatase .

such as

cannot

and 2-deoxyglucose since

than

be feasible.

of oxidativdphosphorylation

is feasible

in

requires

the addition phosphorylation.

the utilization

in the medium

that

not only

glycolysis

or shunt

et -- al. , 1957). As experimental

twelve

to eighteen

genized The

.

homogenates,

It was thought

2-deoxyglucose-6-phosphate

(Wick

fluoride

nucleotides

of hexokinase

glucose

in tissue

1957).

system

This

rates

to include

(Potter,

technique,

but the addition

workable

G. Warnock

glycolytic

of adenine

of enolase

The normal used,

maximal

it is advisable

the fluoride

is routinely

and Laken

19, lg.59 to observe

the bypassing

IN GLYCOLYSIS

Department of Biochemistry University School of Medicine Nashville, Tennessee

‘Vanderbilt

Received

Sept. 19S9

animals hours

in ten volumes

Warburg

flasks

Swiss

before 0.25

strain

mice

the experiment. 0.05

M sucrose,

had the following

additions:

*

Investigator of the Howard Hughes Medical supported by the National Science Foundation 152

were

used,

starved

The

livers

were

M in potassium in the sidearm

Institute. This Grant #5833.

homo-

chloride. 12 FM

research

was

Vol. 1, No. 3

BIOCHEMICAL

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

fructose-di-phosphate and Colowick,

(FDP) 1957).

In the main

diphosphopyridine

nucleotide

sodium

phosphate,

potassium

tinamide,

0.03

of 3.0 ml.

of FDP

experiments. twenty pyruvic lates

cent.

acid without

by paper uptake

#2,

of hexokinase

gave

phosphate Intact

cells, ATP

to storage

forms

introduction

on ATP, directly

Table oxygen

especially connected

After

in,

per

The

identity

uptake

of 2.7

acid

state,

energy

such

by the 2 -deoxyglucose carbohydrate

have active

as creatine system

The

of only

oxygen decrease; 3 units

a formation

sensitive

of

to decreased

is. a continuous

drain

on

transport

and conversion

phosphate

or polyphosphates.

to imitate -hexokinase

metabolism, 153

.

but still

accumu-

was verified

than the pyruvate

is not very uptake

of typical

Pyruvic

the inclusion PM,

on

by about

of the acid

vessel

processes,

of such an acceptor

with

was

(3) It decreases

cent.

system

glycolysis

in the resting

of high

potassium

effect

the result

reproducibly

ninety

I, in another

synthetic

volume

10 minutes

and incubation

I shows

formation.

but the oxygen

even

cent

has a profound

uptake

to the acceptor

(4) The

for

37’C.

lactate

system.

an oxygen

concentrations,

synthesized

The

Table

JLM pyruvate.

in a final

of the 2.4 -dinitrophenylhydrazone

sensitive

in experiment

120 @4 nico-

20 per

and hexokinase

glycolysis.

chromatography

50 JLM

0.2 ml

tipped

1.5 PM

minutes.

by at least

an acceptor

is more

1.29

forty

chloride,

with

(Darrow

included:

ml homogenate

air,

were

(2) It decreases

formation

-triphosphate,

temperature

contents

(1) It accelerates

per

pM adenosine

Bath

the deoxyglucose

the end products

were

c , and 0.3

well.

for an additional

Including

hexokinase

compartment

gas phase

the sidearm

continued

of yeast

20 pM magnesium

pH 7.6,

in the center

equilibration

, 3.0

PM cytochrome

Final

hydroxide

and 15 units

Sept. 1959

the normal couple

institutes

drain

which again

is not a

* 0.58

Experiment

PM

endogenous

-2.42 -0.23

None

-phosphate*

present.

-2.02

-hexokinase

was

t7.20

-2.30

None

phosphate

t5.82

-2.19

-2-deoxyglucose

lactate

t5.60

t4.70

+3.70

+3.40

(PM)

-2.83

A

the end glycolysis

None

02

on

(PM)

A

effect of an acceptor system of aerobic fructose-diphosphate

I

Omission from vessel

The

Table

products

A pyruvate

-0

to.41

-

-

0.37

5.44

+10.41

7.4

9.2

+4.76

t

-

phosphate (PM)

-10.43

&

-0

t5.19

+0.15

(04

Vol. 1, No. 3

BIOCHEMICAL

property tions

of whole of this

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

cells,

system

will

not normally

found

in homogenates.

be published

elsewhere.

Sept. 1959 Applica-

References Darrow,

R.

Potter,

A. and Colowick,

V. R. Stauffer

(1957)

in W.

, Manometric

S. P.

(1957)

unpublished

W. Umbreit,

R.

H.

Techniques,

Burgess

procedure.

Burris, Publishing

and J. F. Co. ,

Minneapolis. Wick,

A. Biol.

N.,

Drury,

Chem.

D. R., , 224,

Nakada,

H.

963 (1957).

155

T. and

Wolfe,

J. B.,

J.