The contact-system dependent plasminogen activator from human plasma: identification and characterization

D J Binnema, G Dooijewaard, J J van Iersel, P N Turion, C Kluft

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Apart from tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA), a third PA appears to occur in human plasma. Its activity is initiated when appropriate triggers of the contact system are added, and the activation depends on the presence of factor XII and prekallikrein in plasma. The activity of this, so-called, contact-system dependent PA accounts for 30% of the PA activity in the dextran sulphate euglobulin fraction of plasma and was shown not to be an intrinsic property of one of the contact-system components, nor could it be inhibited by inhibitory antibodies against t-PA or u-PA. We have succeeded in identifying this third PA in dextran sulphate euglobulin fractions of human plasma. Its smallest unit (SDS-PAGE) is an inactive 110 kDa single-chain polypeptide which upon activation of the contact system is converted to a cleaved, disulphide-bridged molecule with PA activity. The native form, presumably, is an oligomer, since the apparent Mr on gel-chromatography is 600,000. The IEP is 4.8, much lower than that of t-PA and u-PA. Although the active 110 kDa polypeptide cannot be inhibited by anti-u-PA, it yet comprises a 37 kDa piece with some u-PA related antigenic determinants. However, these determinants are in a latent or cryptic form, only detectable after denaturation by SDS. The 110 kDa polypeptide is evidently not a dimer of 55 kDa u-PA or a complex of u-PA with an inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS)

Original languageEnglish
Pages (from-to)390-397
JournalThrombosis and haemostasis
Volume64
Issue number3
Publication statusPublished - 30 Nov 1990

Fingerprint

Forensic Anthropology
Plasminogen Activators
Urokinase-Type Plasminogen Activator
Serum Globulins
Dextran Sulfate
Peptides
Prekallikrein
Factor XII
Plasminogen Inactivators
Tissue Plasminogen Activator
Disulfides
Gel Chromatography
Epitopes
Polyacrylamide Gel Electrophoresis
Antibodies

Keywords

  • Antibodies
  • Blotting, Western
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Precursors
  • Humans
  • Immunoassay
  • Kallikreins
  • Plasminogen Activators
  • Prekallikrein
  • Urokinase-Type Plasminogen Activator
  • Research Support, Non-U.S. Gov't

Cite this

Binnema, D J ; Dooijewaard, G ; van Iersel, J J ; Turion, P N ; Kluft, C. / The contact-system dependent plasminogen activator from human plasma : identification and characterization. In: Thrombosis and haemostasis. 1990 ; Vol. 64, No. 3. pp. 390-397.
@article{39c0d4f1ad174ef3ac91a4c5c125f615,
title = "The contact-system dependent plasminogen activator from human plasma: identification and characterization",
abstract = "Apart from tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA), a third PA appears to occur in human plasma. Its activity is initiated when appropriate triggers of the contact system are added, and the activation depends on the presence of factor XII and prekallikrein in plasma. The activity of this, so-called, contact-system dependent PA accounts for 30{\%} of the PA activity in the dextran sulphate euglobulin fraction of plasma and was shown not to be an intrinsic property of one of the contact-system components, nor could it be inhibited by inhibitory antibodies against t-PA or u-PA. We have succeeded in identifying this third PA in dextran sulphate euglobulin fractions of human plasma. Its smallest unit (SDS-PAGE) is an inactive 110 kDa single-chain polypeptide which upon activation of the contact system is converted to a cleaved, disulphide-bridged molecule with PA activity. The native form, presumably, is an oligomer, since the apparent Mr on gel-chromatography is 600,000. The IEP is 4.8, much lower than that of t-PA and u-PA. Although the active 110 kDa polypeptide cannot be inhibited by anti-u-PA, it yet comprises a 37 kDa piece with some u-PA related antigenic determinants. However, these determinants are in a latent or cryptic form, only detectable after denaturation by SDS. The 110 kDa polypeptide is evidently not a dimer of 55 kDa u-PA or a complex of u-PA with an inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS)",
keywords = "Antibodies, Blotting, Western, Electrophoresis, Polyacrylamide Gel, Enzyme Precursors, Humans, Immunoassay, Kallikreins, Plasminogen Activators, Prekallikrein, Urokinase-Type Plasminogen Activator, Research Support, Non-U.S. Gov't",
author = "Binnema, {D J} and G Dooijewaard and {van Iersel}, {J J} and Turion, {P N} and C Kluft",
year = "1990",
month = "11",
day = "30",
language = "English",
volume = "64",
pages = "390--397",
journal = "Thrombosis and haemostasis",
issn = "0340-6245",
publisher = "Schattauer",
number = "3",

}

Binnema, DJ, Dooijewaard, G, van Iersel, JJ, Turion, PN & Kluft, C 1990, 'The contact-system dependent plasminogen activator from human plasma: identification and characterization', Thrombosis and haemostasis, vol. 64, no. 3, pp. 390-397.

The contact-system dependent plasminogen activator from human plasma : identification and characterization. / Binnema, D J; Dooijewaard, G; van Iersel, J J; Turion, P N; Kluft, C.

In: Thrombosis and haemostasis, Vol. 64, No. 3, 30.11.1990, p. 390-397.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - The contact-system dependent plasminogen activator from human plasma

T2 - identification and characterization

AU - Binnema, D J

AU - Dooijewaard, G

AU - van Iersel, J J

AU - Turion, P N

AU - Kluft, C

PY - 1990/11/30

Y1 - 1990/11/30

N2 - Apart from tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA), a third PA appears to occur in human plasma. Its activity is initiated when appropriate triggers of the contact system are added, and the activation depends on the presence of factor XII and prekallikrein in plasma. The activity of this, so-called, contact-system dependent PA accounts for 30% of the PA activity in the dextran sulphate euglobulin fraction of plasma and was shown not to be an intrinsic property of one of the contact-system components, nor could it be inhibited by inhibitory antibodies against t-PA or u-PA. We have succeeded in identifying this third PA in dextran sulphate euglobulin fractions of human plasma. Its smallest unit (SDS-PAGE) is an inactive 110 kDa single-chain polypeptide which upon activation of the contact system is converted to a cleaved, disulphide-bridged molecule with PA activity. The native form, presumably, is an oligomer, since the apparent Mr on gel-chromatography is 600,000. The IEP is 4.8, much lower than that of t-PA and u-PA. Although the active 110 kDa polypeptide cannot be inhibited by anti-u-PA, it yet comprises a 37 kDa piece with some u-PA related antigenic determinants. However, these determinants are in a latent or cryptic form, only detectable after denaturation by SDS. The 110 kDa polypeptide is evidently not a dimer of 55 kDa u-PA or a complex of u-PA with an inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS)

AB - Apart from tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA), a third PA appears to occur in human plasma. Its activity is initiated when appropriate triggers of the contact system are added, and the activation depends on the presence of factor XII and prekallikrein in plasma. The activity of this, so-called, contact-system dependent PA accounts for 30% of the PA activity in the dextran sulphate euglobulin fraction of plasma and was shown not to be an intrinsic property of one of the contact-system components, nor could it be inhibited by inhibitory antibodies against t-PA or u-PA. We have succeeded in identifying this third PA in dextran sulphate euglobulin fractions of human plasma. Its smallest unit (SDS-PAGE) is an inactive 110 kDa single-chain polypeptide which upon activation of the contact system is converted to a cleaved, disulphide-bridged molecule with PA activity. The native form, presumably, is an oligomer, since the apparent Mr on gel-chromatography is 600,000. The IEP is 4.8, much lower than that of t-PA and u-PA. Although the active 110 kDa polypeptide cannot be inhibited by anti-u-PA, it yet comprises a 37 kDa piece with some u-PA related antigenic determinants. However, these determinants are in a latent or cryptic form, only detectable after denaturation by SDS. The 110 kDa polypeptide is evidently not a dimer of 55 kDa u-PA or a complex of u-PA with an inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS)

KW - Antibodies

KW - Blotting, Western

KW - Electrophoresis, Polyacrylamide Gel

KW - Enzyme Precursors

KW - Humans

KW - Immunoassay

KW - Kallikreins

KW - Plasminogen Activators

KW - Prekallikrein

KW - Urokinase-Type Plasminogen Activator

KW - Research Support, Non-U.S. Gov't

M3 - Article

C2 - 2128968

VL - 64

SP - 390

EP - 397

JO - Thrombosis and haemostasis

JF - Thrombosis and haemostasis

SN - 0340-6245

IS - 3

ER -