Lactate dehydrogenase isoenzymes in human hearts having decreased oxygen supply

BIOCHEMICAL

Vol. 33, No. 3, 1968

AND BIOPHYSICAL

LACTATE DEHYDROGMASB ISOENZYMES DBCRUSED

HAVING

Joseph

M. Ballo,

(Tufts)

M. D. and Joseph

and Mallory

Institute

IN HUM&I HURTS

CKYGEN SUPPLY

Messer,

V.

RESEARCH COMMUNICATIONS

H. D.,

of Pathology,

Circulation

Boston

City

Laboratory

Hospital,

Boston,

Masaachuaetts.

Received October 2, 1968 In most

animal

tains

two polypeptide

cular

weight,

toward

higher

It

xation

of anaerobic relatively

At

bition.

This amount

Heart

suggested

that

of substrate

inhibition

between ates

08613,

advanced

the

of pyruvate

1961;

two subunits.

(isoenzymes

H,,,, H3M, H2kf2,

normal

having

muscle hearts

infants

morphologic

mass showed

or thoae revealed

mechanism

available

for

with

a either

a similar enhanced

patutili-

pathways. concentrations,

of the

inhibition

related

NAU4 , pyruvate,

have

centrations

is

these

and increased

chain

H

is

to the and LDH

hypothesis may assure

LDH exhibits (predominant

directly

of H chains in the enzyme (Dawson at al, this

behavior

et al,

from human hearts

of newborn

in mole-

of H and M chains

is an adaptive

pyruvate

a property

of

con-

differing

Kaplan

tetramers

LDH from

tissue this

1960;

percentage

perfusion

glycolytic high

is

possible

than

al,

1.1.1.27)

and kinetic

is a tetramer

dehydrogenase

of M chains

tern.

et

relative

tissue

alone. is

five

by the

of diminished proportion

enzyme

the

Lactate

characteristic

the

of

mobility

(Raplan

The intact

(LDH B.C.

H and H subunits,

electrophoretic

determined

combination.

dehydrogcnase

designated

and cofactors

proportion

HE+, Mq) is

evidence

chains,

1962).

al,

The tissue

for

lactate

antigenicity,

substrates

Cahn et

tissues,

formation (Gutfreund that

to

et al,

inhibition

This work was suppprted by U.S.P.H.S. grants and by a Research Grant fran the Medical

inhi-

muscle),

and

percentage

the

It has been documented that

of an abortive

a constaet

487

in heart

proportional

1964).

substrate

flow

terniary

1968).

Raplan

of LDH

by high

of

substrate

EIB-5687, Foundation,

complex and associtissue into

HB-8719, Boston.

conpathways

and H& Mass-

Vol. 33, No. 3, 1968

of oxidative which

phosphorylation,

rely

on oxidative

predominantly tal

relying

gators

have

inhibited)

the

support

the

major

isoeneyme,

while

of

this

such

source

for

as the

heart,

of energy,

tissues

glycolysis inhibited)

validity

Kaplan

tissues,

their

on anaerobic

RESEARCH COMMUNICATIONS

that

for

of M4 (non-pyruvate

questioned

Cur findings

observed

phosphorylation

partially

a predominance

AND BIOPHYSICAL

having

H4 (pyruvate

muscle,

have

BIOCHEMICAL

such

energy

have

as skele-

production,

isoensyme.

Some investi-

hypothesis

(Vesell

were

from

and Pool,

1966).

hypothesis. METHODS

Sections cular

free

buffer

of hearts wall

and frosen.

at pli 7.4.

the

rate

buffer

at

sodium

pyruvate.

at autopsy

Homogenates

were

Electrophoretograms

in 30 n&i phosphate uring

obtained

buffer,

pli 7.0,

of disappearance

37OC) at

low

were at

x 10e4M)

prepared

3OC.

the

acetate

was assayed

340 n&l (pH 7.5, (3.3

ventri-

on cellulose

LDH activity

and high

left

in 100 ml4 phosphate

performed

of NADH at

(3.3

taken

by meas-

100 m?4 phosphate

x lO’3M)

concentration

of

REWLTS

As estimated centage

of M chains

combination denced

males).

Those

(Group

concentrations these

with I)

than

IV)

patterns

relevant

contained

greater only

Total

activity

increased

and high/low

M

than

one

(Group

II)

of LDH (in

subunits,

pyruvats

kinetics.

488

from

as evior old

325 gm for

females,

in Table

or neither

pyruvate (Group

tissue)

III) was not

950 to 2600 gm newborns

as evidenced

I.

and cardiac

at high

units/gm

a

or recent

tissue

LDH activity

per-

having

are presented

perfused

Hearts

among groups.

patients

fibrosis,

data

of poorly

an increased

enlargement,

(greater

morphologic

having

those

cardiac

weight

had relatively

different also

heart

I),

and cardiac

focal

a combination

characteristics.

significantly

total

hearts

LDH from

perfusion

narrowing,

Other

hearts

hypertrophy

phoretic

artery

(Figure

in cardiac

tissue

and increased

375 gm for

electrophoretograms

was present

by coronary

(Group

the

of diminished

infarction,

of

from

by elactro-

Vol. 33, No. 3, 1968

BlOCHEMlCAL

AND BIOPHYSICAL

RESEARCH COMMUNICATIONS

Figure 1. Cellulose acetate LDH electrophoretograw of heart homogeProp left to right the banda represent isoenaymea I&, M3H, H2H2, &iH3, natea. The heavy double line is the application point; the anode (-) ir end I+,. to the right. (1) and (2) Coronary artery disease in association with cardiac enlargement (Group I), showing increased concentrations of M4 and M3H ahowiag normal pattern (Group iaoanaymea; (3) Coronary artery disease alone, 11) ; (4) Neither cardiac enlargement nor coronery artery disease (Group III), showing normel pattern; (5) 1700 gm newborn (Group IV), showing pronounced M-chain shift in LDH iaoenayma distribution.

DISCUSSION Batradiol ment

all

exhibit

stimulation enhanced

of rates

the

uterus, of protein

489

neoplaaia, synthesis,

and embryonic and alterations

davelopin LDH

(%)-

Wall

Wall

Thicknerr

Thickness (cm)

(cm)

M~ROL~IC,

+, .04

+ .lO

40.1 -p

L--p

2 3.7

35 + 10

.20

2.0

496 +, 46

60+6

5/3

8

I

AND BIOCBgMICAL

2 .02

2 .lO

- .015-

29.9 = .OvLp

fr 2.0

15 2 5

.18

1.50

399 t 30

70 2 4

6/7

13

II

Group*

+, .05

30 2 1.1

15 f 5

.15 +, .03

1.40

330 $23

55 +, 9

3/l

4

III

OP PATIENT

Patient

CBARACTgRIzATICN

I

-

w-w

-MS

s-s

w-w

4/l

5

IV

2 4.3

emall

corrected

aample

(3.3x10S3&f)

diseaoe pyruvate

coronary artery IV. Newborns.

.025-

42.8

45 +, 10

GRUJFS

Coronary artery dFaease with cardiac enlargement (hypertrophy); If. Either enlargement; III. Neither coronary artery direaoe nor cardiac enlargement; M-type isoentyme present estimated fran electrophoretograms. high/low ir the relative activity (in %) of LDH at high concentrations of to that at low concentrations (3.3~10%). population mean8 + standard error of the mean and p value6 are derived from

high/low

Activity

* Group 1. or cardiac * Percent Activity aa compared Valuer are rtatisticr.

M Chain8*

Percent

Ventricular

Ventricular

Left

Right

Weight

Reart

Age (years)

(gm)

of Cases

Hale haale

Number

CLINICAL,

Table

Vol. 33, No. 3, 1968

isoentyme

composition

Goldman,

et

muscles,

Goldberg

increases rats

al,

1964;

exposed

servations

perfusion

to

8. 9. 10. 11.

al, It

a necessary

oxygen

1968), is

delayed

in

capacity

This

adaptation to total the

The relative

may reflect

in cardiac

LDH

a similar

while of

newborn the

with that

adult

our

ob-

cardiac rat.

an adaptive

a combination

of

Adult

the adult

with

glycolysis

hamstring involved

achievement

however,

1964;

1968).

consistent

factor

enlargement.

rat

of M chains,

of interest,

production.

tension

a shift

findings

associated

increased

with

hypertrophy

as demonstrating

of anaerobic

lactate

proportion shwed

et al,

(Goldberg,

shwed

co-existing

composition

by the

for lw

et

experiments

proteins

a higher

(Goodfriend

work-induced

tensions

conditions

and cardiac

evidenced

tissue

oxygen

state

In

compensatory

(Wager

contribution

in each

1963).

may be interpreted

LDH isoenzyme

tissue

found

al,

human heart.

was not

heart

6. 7.

lw

pattern

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

and contractile

similar

in the

enhanced

2. 3. 4. 5.

that

tward

Our findings

1.

et

soluble

under

enlargement

merits,

Fine

distribution

reared

tissue

been

to chronic

H-predominant

heart

have

found

in both

isoensyme rats

BIOCHEMICAL

of

change

in

decreased

may underlie heart

chronically resistance

energy

the require-

under-perfused of

fetal

heart

mechanism.

Cahn, R.D., Kaplan, N.O., Levine, L., and Zwilling, E., Science 136, 962 (1962). Dawaon, D.M., Goodfriend, T.L., and Kaplan, N.O., Science 143, 929 (1964). Fine, I.H., Kaplan, N.O., and Kuftinec, D., Biochemistry 2, 116 (1963). Goldberg, A.L., J. Cell Biol. 36, 653 (1968). N.O., and &all, T.C., Cancer Research 24, 389 Goldman, R.D., Kaplan, (1964) . Goodfriend, T.L., and Kaplan, N.O., J. Biol. Chem. 239, 130 (1964). Gutfreund, H., Cantwell, R., McMurry, C.H., Griddle, R.S., and Hathaway, G Biochem. J. lG6, 683 (1968). K$an, N.O., Ciotti, H.M., Ramolsky, M., and Bieber, R.E., Science 131, 392 (1960). M., Ann. N.Y. Acad. Sci. 94, 701 (1961). Kapl8n, W.O., and Ciotti, Mager, H., Blatt, W.P., Natale, P.J., and Blatteis, C.W., Am. J. Physiol. 23.5, 8 (1968). Vesell, S.S., and Pool, P.E., Proc. Nat. Acad. Sci. (U.S.) 55, 756 (1966).

491