THROMBOSIS RESEARCH Printed in the

INHIBITION

OF

Pp,rgamon

States

C OX31UNI

BRIEF PURIFIED

PLASMA AND

Pp. 223-225,

7,

Vol.

United

CAT1

KALLIKREIN

Press,

197j

Inc.

OX

BY ANTITHROMBIN

III

HEPARIN

Anne Marie Vennertjd and Knut LaakeX Department

of Pharmacology, University

(Received

to Yin et al.

identical

with the heparin of heparin

(1) antithrombin cofactor

antithrombin

factor Xa and the factor XIa heparin

forms a complex

serine proteinases

antithrombin

importance trinsic (5).

III of human plasma

III inhibits

(2). Rosenberg

in and related

thrombin,

III which

is rapidly

of heparin

is

Xa. In the the

(3) has proposed

that

inactivates

to the coagulation

that plasmin

III in the presence

Recently,

form 9.5.1974. Godal)

and the antifactor

with antithrombin

and it has been observed

of Pharmacy,

of Oslo, Norway

in revised 29.4.1975; Accepted by Editor H.C.

According

presence

Institute

cascade,

inactivated

by

(4).

plasma

kallikrein

has been reported

in the initial

activation

of the factor XII in the in-

blood coagulation

Plasma

kallikrein

and the factor link between

IX

also activates

(7). Plasma

the initial

XPresent

system by negatively

Department

kallikrein

Haematological

IX, University

surfaces

plasminogen

may also represent

parts of the intrinsic

address:

Oslo, Norway.

purified

charged

to be of

a

and the extrinsic

Research

Clinic,

(6)

Laboratory,

Ullevaal

Hospital,

blood

coagulation

systems

arin and antithrombin factor VII purified

III somehow

(9). The present

plasma

(a), and it has been observed

kallikrein

interfere

experiments

that hep-

with the activated

were undertaken

is inactivated

to see if

by antithrombin

III and

heparin. Partially activated viously

purified

with purified

human plasma

factor XII fragments

shown that the activity

a-N-benzoyl-L-arginine

prekallikrein

was

(11). We have pre-

of such preparations

ethyl ester

(10)

towards

(BAEe) is due to the activat-

05-

Fig. 1 Inhibition of plasma kallikrein by antithrombin III and heparin. -c: Plasma kallikrein, antithrombin III and heparin _ : Plasma kallikrein and antithrombin III & : Plasma kallikrein and heparin + : Plasma kallikrein alone The final concentrations in the incubations were: Plasma kallikrein 0.7 BAEe U/ml, antithrombin III 0.4 U/ml (defining 1 U as the amount present in 1 ml titrated plasma), and heparin 0.85 U/ml (obtained dry from A/S Apothekernes Laboratorium for Specialpreparatgr, Oslo). All incubations were carried out at 25 C in 0.05 M Tris/HCl pH 7.4. The BAEe esterase activity in the incubations was determined at intervals with NAD and alcohol dehydrogenase (10).

IMLLIKREIN

t’cJl.~,lio.l

ion of plasma

L AXTITHROMBIS

Antithrombin

16).

prekallikrein

from human titrated

plasma by a two-step

inity chromatography

on heparin-coupled

DEAE-Sephadex

A-50 chromatography

No inhibition observed

ever, the kallikrein and antithrombin

plasma

antithrombin

plasma

kallikrein

a2-macroglobulin that plasma vation

kallikrein

of the initial

the results impaired heparin

probably

intrinsic

sustaining

the results

Plasma pathogenesis

ological

ment with heparin

system,

of factor XII is also during

of factor VII by the plasma

kallikrein

and antithrombin

angioneurotic of plasma

III,

(13).

edema which

(14). The rapid III and heparin

concentrations

might be of benefit

acti-

to play a role in the

by antithrombin

and pharmacological

of

and the

coagulation

experiments

is supposed

of the hereditary

kallikrein

inhibitor

for the normal

through

by heparin

of previous

kallikrein

blood and

in vitro and possibly

system mediated

to lack of the Ci-inactivator ion of plasma

is necessary

in vivo. The activation

also counteracted

(Fig. 1).

As it is now accepted

mean that the activation

coagulation

is probably

acting

stages of the intrinsic

plasma

III. How-

in the incubations

of proteinases.

was

when both heparin

to the Ci-inactivator

activity

in heparinized treatment

or antithrombin

III may be a rapidly

inhibitor

aff-

gel and

activity

that in heparinized

in addition

employing

polyacrylamide

inactivated

III were present indicate

procedure

kallikrein

with heparin

was rapidly

The results

III was purified

(12).

of the plasma

upon incubation

225

III

indicates

is due inactivatin physi-

that treat-

to such patients.

REFERENCES 1. YIN, E.T., WESSLER, S., and STOLL, P.J. Identity of plasma-activated factor X inhibitor with antithrombin III and heparin cofactor. J. biol. Chem.: 246, 3712-3719, 1971.

2. DAWS, P.S., HICKS, M., and ROSENBERG, R.D. Anticoagulant action of heparin. Nature: 246, 355-357, 1973. 3.

ROSENBERG, R.D. Actions and interactions of antithrombin and heparin. N. Eng. J. Med.: 292, 146-151, 1975.

4. HIGHSMITH, R.F. and ROSENBERG, R.D. The inhibition of human plasmin by antithrombin-heparin cofactor. J. biol. Chem.: 249, 4335-4338, 1974. -5.

COCHRANE, C.G., REVAK, S.D., and WJEPPER, K.D. Activation of Hageman factor in solid and fluid phases. J. exp. : 138, 1564-1538, 1973. Med. -6. LAAXE, K. and VENNERgD, A.M. Factor XII-induced fibrinolysis: Studies on the separation of prekallikrein, plasminogen proactivator, and factor XI in human plasma. Thromb. Res.: Q, 285-302, 1974. B., LAAKE, 7. %TERUD, factor IX. In press

K., and PRYDZ, H. Activation in: Thromb. Diath. haemorrh.

Of

8. GJijNNAESS, H. Cold promoted activation of factor VII. II. Identification of the activator. Thromb. Diath. haemorrh.: 28, 169-181, 1972. 9.

GODAL, H-C., RYGH, M., and LAAKE, K. Progressive inactivation of purified factor VII by heparin and antithrombin 111. Thromb. Res.: 2, 773-775, 1974.

10.

LAAKE, K. and VENNERGD, A.M. Determination of factor XII in human plasma with arginine proesterase (prekallikrein). I. Preparation and properties of the substrate. Thromb. Res.: 2_, 393-407, 1973.

11. VENNERijD, A.M. and LAAKE, K. Isolation and characterization of a prealbumin activator of prekallikrein from acetone-activated human plasma. Thromb. Res.: 4, 103-108, 1974. 12. MILLER-ANDERSON, M., BORG, H., and ANDERSSON, L.-O. Purification of antithrombin III by affinity chromatography. Thromb. Res.: 2, 439-452, 1974. 13. GODAL, H-C., GRAVEM, K., and LAAKE, K. Progressive inactivation of factor VII in the cold by heparin. Thromb. Res.: fi, 707-710, 1974. 14. DENNEHY, J.J. Hereditary angioneurotic edema. Report of a large kindred with defect in C'l esterase inhibitor and review of the literature. Ann. intern. Med.: 73, 55-59, 1970.

Inhibition of purified plasma kallikrein by antithrombin III and heparin.

THROMBOSIS RESEARCH Printed in the INHIBITION OF Pp,rgamon States C OX31UNI BRIEF PURIFIED PLASMA AND Pp. 223-225, 7, Vol. United CAT1 KA...
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