Download PDF
Thromb Haemost 1982; 48(03): 320-324
DOI: 10.1055/s-0038-1657290
Original Article
Schattauer GmbH Stuttgart

The Suitability of Various Plasmin Preparations for the Functional Assay of α2-Antiplasmin in Plasma

C Kluft
The Gaubius Institute, Health Research Division TNO, Leiden, The Netherlands
,
D W Traas
The Gaubius Institute, Health Research Division TNO, Leiden, The Netherlands
,
A F H Jie
The Gaubius Institute, Health Research Division TNO, Leiden, The Netherlands
,
E Hoegee-de Nobel
The Gaubius Institute, Health Research Division TNO, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 24 May 1982

Accepted 08 November 1982

Publication Date:
13 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Various plasmin preparations were tested for their suitability for use in the assay of α2-antiplasmin in blood plasma by the immediate plasmin inhibition test.

Activation of plasminogen, viz., 1-Glu-plasminogen and/or 77-Lys-plasminogen, by immobilized urokinase results in plasmin preparations suitable for this α2-antiplasmin test. Plasmins obtained by »spontaneous« activation procedures in glycerol containing solutions, however, appeared not to be suitable. In this second group, the behaviour of the plasmins resembles that of 442-Val-plasmin (miniplasmin), which is known to show a low inactivation rate with α2-antiplasmin due to the absence of lysine-binding sites in the plasmin molecule.

Evidence is presented that, in the nonsuitable plasmins, the lysine-binding sites, although not completely absent, have at least partly lost their functional integrity.

Keywords

Fibrinolysis - Plasmin - α2-Antiplasmin - H-D-Val-Leu-Lys-pNA
 

  • References

  • 1 Gallimore HJ, Amundsen E, Aasen AO, Larsbraaten M, Lyngaas K, Svendsen L. Studies on plasma antiplasmin activity using a new plasmin specific chromogenic tripeptide substrate. Thromb Res 1979; 14: 51-60
    CrossrefPubMedGoogle Scholar
  • 2 Teger-Nilsson AC, Friberger P, Gyzander E. Determination of a new rapid plasmin inhibitor in human blood by means of a plasmin specific tripeptide substrate. Scand J Clin Lab Invest 1977; 37: 403-409
    CrossrefPubMedGoogle Scholar
  • 3 Edy J, De Cock F, Collen D. Inhibition of plasmin by normal and antiplasmin-depleted human plasma. Thromb Res 1976; 8: 513-518
    CrossrefPubMedGoogle Scholar
  • 4 Kluft C, Vellenga E, Brommer EJP. Homozygous α2-antiplasmin deficiency. Lancet 1979; 2: 206
    PubMedGoogle Scholar
  • 5 Kluft C, Vellenga E, Brommer EJP, Wijngaards G. A familial haemorrhagic diathesis in a dutch family: an inherited deficiency of α2-antiplasmin. Blood 1982; 59: 1169-1180
    PubMedGoogle Scholar
  • 6 Christensen U, Clemmensen I. Kinetic properties of the primary inhibitor of plasmin from human plasma. Biochem J 1977; 163: 389-391
    CrossrefPubMedGoogle Scholar
  • 7 Wiman B, Collen D. On the kinetics of the reaction between human antiplasmin and plasmin. Eur J Biochem 1978; 84: 573-578
    CrossrefPubMedGoogle Scholar
  • 8 Wiman B, Boman L, Collen D. On the kinetics of the reaction between human antiplasmin and a low-molecular-weight form of plasmin. Eur J. Biochem 1978; 87: 143-146
    CrossrefPubMedGoogle Scholar
  • 9 Christensen U, Sottrup-Jensen L, Magnusson S, Petersen TE, Clemmensen I. Enzymic properties of the neo-plasmin-Val-442 (miniplasmin). Biochim Biophys Acta 1979; 567: 472-481
    CrossrefPubMedGoogle Scholar
  • 10 Philo RD, Gaffney PJ. Plasmin potency estimates: influence of the substrate used in assay. Thromb Haemostas 1981; 45: 107-109
    PubMedGoogle Scholar
  • 11 Kluft C, Brakman P, Veldhuyzen-Stolk EC. Screening of fibrinolytic activity in plasma euglobulin fractions on the fibrin plate: Progress in Chemical Fibrinolysis and Thrombolysis. Davidson JF, Samama MM, Desnoyers PC. (Eds) Raven Press; New York: 1976. 2 57-65
    Google Scholar
  • 12 Summaria L, Spitz F, Arzadon L, Boreiska JG, Robbins KC. Isolation and characterization of the affinity chromatography forms of human Glu- and Lys-plasminogens and piasmins. J Biol Chem 1976; 251: 3693-3699
    PubMedGoogle Scholar
  • 13 Bethell GS, Ayers JS, Hancock WS, Hearn MTW. A novel method of activation of cross-linked agaroses with 1,1’-carbonyldiimidazole which gives a matrix for affinity chromatography devoid of additional charged groups. J Biol Chem 1979; 254: 2572-2574
    PubMedGoogle Scholar
  • 14 Haverkate F, Brakman P. Fibrin plate assay: Progress in Chemical Fibrinolysis and Thrombolysis. Davidson JF, Samama MM, Desnoyers PC. (Eds) Raven Press; New York: 1971. 1 151-159
    Google Scholar
  • 15 Chase Jr T, Shaw E. Comparison of the esterase activities of trypsin, plasmin and thrombin on guanidinebenzoate esters. Titration of enzymes. Biochemistry 1969; 8: 2212-2224
    CrossrefPubMedGoogle Scholar
  • 16 Sottrup-Jensen L, Claeys H, Zajdel M, Petersen TE, Magnusson S. The primary structure of human plasminogen: I. Isolation of two lysine-binding fragments and one “mini”-plasminogen (MW 38,000) by elastase-catalyzed-specific limited proteolysis: Progress in Chemical Fibrinolysis and Thrombolysis. Davidson JF, Rowan RM, Samama MM, Desnoyers PC. (Eds) Raven Press; New York: 1978. 3 191-209
    Google Scholar
  • 17 Kirkwood TBL, Campbell PJ, Gaffney PJ. A standard for human plasmin. Thrombos Diathes Haemorrh 1975; 34: 20-31
    PubMedGoogle Scholar
  • 18 Sgouris JT, Inman JK, McCall KB, Hyndman LA, Anderson HD. The preparation of human fibrinolysin (plasmin). Vox Sanguinis 1960; 5: 357-376
    CrossrefPubMedGoogle Scholar
  • 19 Deutsch DG, Mertz ET. Plasminogen: purification from human plasma by affinity chromatography. Science 1970; 170: 1095-1096
    CrossrefPubMedGoogle Scholar
  • 20 Alkjaersig N, Fletcher AP, Sherry S. The activation of human plasminogen. I. Spontaneous activation in glycerol. J Biol Chem 1958; 233: 81-85
    PubMedGoogle Scholar
  • 21 Allen RA. The effect of poly-lysine on the activation of plasminogen: Progress in Fibrinolysis. Davidson JF, Nilsson IM, Åstedt B. (Eds) Churchill Livingstone; Edinburgh: 1981. 5 134-138
    Google Scholar
  • 22 Claeson G, Aurell L, Karlsson G, Friberger P. Substrate structure and activity relationships: Progress in Chemical Fibrinolysis and Thrombolysis. Davidson JF, Rowan RM, Samama MM, Desnoyers PC. (Eds) Raven Press; New York: 1978. 3 299-304
    Google Scholar
  • 23 Wohl RC, Summaria L, Robbins KC. Physiological activation of the human fibrinolytic system. Isolation and characterization of human plasminogen variants, Chicago I and Chicago II. J Biol Chem 1979; 254: 9063-9069
    PubMedGoogle Scholar
  • 24 Morris JP, Blatt S, Powell JR, Strickland DK, Castellino FJ. Role of lysine binding regions in the kinetic properties of human plasmin. Biochemistry 1981; 20: 4811-4816
    CrossrefPubMedGoogle Scholar
  • 25 Wohl RC, Summaria L, Robbins KC. Kinetics of activation of human plasminogen by different activator species at pH 7.4 and 37° C. J Biol Chem 1980; 255: 2005-2013
    PubMedGoogle Scholar
  • 26 Lämmle B, Duckert F. Different assessment of plasmin with different substrates. In vitro alteration of plasmin, influence of epsilon-aminocaproic acid and tranexamic acid upon its activity. Thromb Haemostas 1980; 43: 112-117
    PubMedGoogle Scholar
  • 27 Summaria L, Robbins KC. Isolation of a human plasmin derived, functionally active, light (B) chain capable of forming with streptokinase an equimolar light (B) chain. Streptokinase complex with plasminogen activator activity. J Biol Chem 1976; 251: 5810-5813
    PubMedGoogle Scholar
  • 28 Thorsen S, Müllertz S. Rate of activation and electrophoretic mobility of unmodified and partially degraded plasminogen. Effects of 6-amino hexanoic acid and related compounds. Scand J Clin Lab Invest 1974; 34: 167-176
    CrossrefPubMedGoogle Scholar
  • 29 Robbins KC, Summaria L. Purification of human plasminogen and plasmin by gelfiltration on Sephadex and chromatography on diethyl amino ethyl-Sephadex. J Biol Chem 1963; 238: 952-962
    PubMedGoogle Scholar
  • 30 Reddy KNN, Wagner CJ. Differential autolysis of human and canine plasmins. Biochem Biophys Res Commun 1980; 92: 1016-1022
    CrossrefPubMedGoogle Scholar
  • 31 Gaffney PJ, Miller-Andersson M, Kirkwood TBL. Unreliability of chromogenic substrates for assay of the clotting activity of thrombin. Haemostasis 1978; 7: 109-112
    PubMedGoogle Scholar
  • 32 Wiman B, Lijnen HR, Collen D. On the specific interaction between the lysine-binding sites in plasmin and complementary sites in α2-antiplasmin and in fibrinogen. Biochim Biophys Acta 1979; 579: 142-154
    CrossrefPubMedGoogle Scholar
  • 33 Lerch PG, Rickli EE. Studies on the chemical nature of lysine-binding sites and on their localization in human plasminogen. Biochim Biophys Acta 1980; 625: 374-378
    CrossrefPubMedGoogle Scholar
  • 34 Bello J, Bello HR. Chemical modification and cross-linking of proteins by impurities in glycerol. Arch Biochem Biophys 1976; 172: 608-610
    CrossrefPubMedGoogle Scholar
  • 35 Gaffney PJ, Philo RD. Measurements of some haemostatic components using biological and amidolytic assays: Progress in Fibrinolysis. Davidson JF, Nilsson IM, Åstedt B. (Eds) Churchill Livingstone; Edinburgh: 1981. 5 196-202
    Google Scholar