Familial Variant of Antithrombin III (AT III Bligny, 47Arg to His) Associated with Protein C Deficiency

M Wolf; C Boyer-Neumann; P Molho-Sabatier; C Neumann; D Meyer; M J Larrieu

doi:10.1055/s-0038-1645197

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1990; 63(02): 215-219
DOI: 10.1055/s-0038-1645197

Original Article

Schattauer GmbH Stuttgart

Familial Variant of Antithrombin III (AT III Bligny, 47Arg to His) Associated with Protein C Deficiency

M Wolf

^*The Laboratoire d’Hématologie and INSERM U 143, Briis-sour-Forges, France

,

C Boyer-Neumann

^*The Laboratoire d’Hématologie and INSERM U 143, Briis-sour-Forges, France

,

P Molho-Sabatier

^**Hôpital de Bicêtre, Le Kremlin-Bicêtre, the Centre de Recherche Claude Bernard sur les maladies vasculaires périphériques, Briis-sous-Forges, France

,

C Neumann

^***Hopital Broussais, Paris, and the CMC de Bligny, Briis-sous-Forges, France

,

D Meyer

^*The Laboratoire d’Hématologie and INSERM U 143, Briis-sour-Forges, France

,

M J Larrieu

^*The Laboratoire d’Hématologie and INSERM U 143, Briis-sour-Forges, France

› Institutsangaben

Abstract

als PDF herunterladen

Summary

The association of a variant of antithrombin III (AT III Bligny) and protein C deficiency is described in a 36-year-old patient having suffered from severe thrombotic episodes. His mother has protein C deficiency and showed a single episode of thrombophlebitis following surgery. His father, sister and daughter have the variant AT III and are asymptomatic. The abnormal AT III was characterized in plasma by the discrepancy between a normal progressive activity and a reduced heparin cofactor activity. This variant AT III was purified, separated from the normal protein by heparin-Sepharose chromatography and was eluted with increased NaCI concentrations. At pH 7.4, the variant AT III eluted at lower (0.3 to 0.5 M) NaCI concentrations than normal (1 to 1.5 M) AT III, thus demonstrating a decreased affinity for heparin. At pH 6.0, however, the abnormal molecule bound more avidly to heparin-Sepharose and was eluted like normal AT III at pH 7.4. Similarly, the heparin enhancement of intrinsic fluorescence of the variant AT III, markedly reduced at pH 7.4, was normalized at pH 6.0. The abnormal AT III showed a normal antiprotease activity, a normal molecular weight by SDS-PAGE, but displayed only a partial immunological identity with the normal protein. Analysis of amplified genomic DNA from this patient by dot-blot demonstrates a heterozygous substitution of arginine by histidine at position 47.

PDF (389 kb)

Referenzen

References
1 Carrell RW, Christey PB, Boswell DR. Serpins: antithrombin and other inhibitors of coagulation and fibrinolysis, evidence from amino acid sequences. Proceedings of Thrombosis XI^th Congresss Haemostasis Verstraete M, Vermylen J, Lijnen R, Arnout J. eds Leuven University Press; Leuven: 1987: 1-15
2 Marlar RA, Kleiss AJ, Griffin JH. Mechanism of action of human activated protein C, a thrombin-dependent anticoagulant enzyme. Blood 1982; 59: 1067-1072
3 Thaler E, Lechner K. Antithrombin deficiency and thromboembolism. Clin Haematol 1981; 10: 369-390
4 Rosenberg RD, Bauer KA. Thrombosis in inherited deficiencies of antithrombin, protein C and protein S. Hum Pathol 1987; 18: 253-261
5 Owen MC, Borg JY, Soria C, Soria J, Caen J, Carrell RW. Heparin binding defect in a new antithrombin III variant: Rouen, 47 Arg to His. Blood 1987; 69: 1275-1279
6 Fenton JW, Fasco MJ, Stackrom AB. Human thrombins. Production, evalution and properties of α-thrombin. J Biol Chem 1977 252. 3587-3599
7 Wolf M, Boyer C, Lavergne JM, Larrieu MJ. A new familial variant of antithrombin III: “Antithrombin III Paris”. Br J Haematol 1982; 51: 285-295
8 Laurell CB. Quantitative estimation of proteins by electrophoresis in agarose gels containing antibodies. Anal Biochem 1966; 15: 45-52
9 Abildgaard U, Larsen ML. Assay of dermatan sulfate cofactor (heparin cofactor II) activity in human plasma. Thromb Res 1984; 35: 257-266
10 Friberger P, Knos M, Gustavesson S, Aufell L, Claeson G. Methods for determination of plasmin antiplasmin and plasminogen by S-2251. Haemostasis 1978; 7: 138-145
11 Amiral J, Adam M, Plassart V, Minard S, Vissac AM. Immunoassays for the measurement of protein S. In Fibrinolysis: Current Prospects. Gaffney PJ, Castellino FJ, Plow EF, Takada A. eds Libbey and Co; London: 1988: 125-128
12 Boyer C, Rothschild C, Wolf M, Amiral J, Meyer D, Larrieu MJ. A new method for the estimation of protein C by ELISA. Thromb Res 1984; 36: 579-589
13 Francis RB, Patch MJ. A functional assay for protein C in human plasma. Thromb Res 1983; 32: 605-613
14 Nicham F, Guichaoua JF, Contant G, Martinoli JL. Dosage rapide de l’activité de la proteine C. Ann Biol Clin 1988; 46: 805-809
15 Wolf M, Boyer C, Tripodi A, Meyer D, Larrieu MJ, Mannucci PM. Antithrombin III Milano: A new variant with monomeric and dimericinactive antithrombin III. Blood 1985; 65: 496-500
16 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 277: 680-685
17 Nordenman B, Danielsson A, Bjork L. The binding of low-affinity and high-affinity heparin to antithrombin. Eur J Biochem 1978; 90: 1-6
18 Bell GI, Karam JH, Rutter WJ. Polymorphic DNA region adjacent to the 5’ end of the human insulin gene. Proc Natl Acad Sci USA 1981; 78: 5759-5763
19 Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487-494
20 Molho-Sabatier P, Aiach M, Gaillard I, Fiessinger JN, Fischer AM, Chadeuf G, Clauser E. Molecular characterization of seven AT III variants using PCR. Identification of a new mutation : 384 Ala-Pro. J Clin Invest 1989 84. in press
21 Koide T, Takahashi K, Odani S, Ono T, Sakuragawa N. Isolation and characterization of a hereditary abnormal antithrombin: antithrombin III Toyama. Thromb Res 1983; 31: 319-328
22 Fischer AM, Gazengel C, Dautzenberg MD, Benhammou J, Beguin S, Vergoz D. A new case of abnormal antithrombin III (AT III): biological characteristics and thromboembolic accidents management. Thromb Haemostas 1983; 50: 358
23 Fischer AM, Cornu C, Sternberg C, Meriane F, Dautzenberg MD, Chafa O, Beguin S, Desnos M. Antithrombin III Alger: a new homozygous AT III variant. Thromb Haemostas 1986; 55: 218-221
24 Boyer C, Wolf M, Vedrenne J, Meyer D, Larrieu MJ. Homozygous variant of antithrombin III: AT III Fontainebleau. Thromb Haemostas 1986; 56: 18-22
25 Moerloose PA, Reber G, Vernet Ph, Minazio Ph, Bouvier CA. Antithrombin III Geneva: a hereditary abnormal AT III with defective heparin cofactor activity. Thromb Haemostas 1987; 57: 154-157
26 Duchange N, Chasse JF, Cohen GN, Zakin MM. Molocular characterization of the antithrombin III Tours deficiency. Thromb Res 1987; 45: 115-121
27 Chang JY, Tran TH. Antithrombin III Basel: identification of a proleu substitution in a hereditary abnormal antithrombin with impaired heparin cofactor activity. J Biol Chem 1986; 261: 1174-1176
28 Aiach M, Francois D, Priollet P, Capron L, Roncato M, Alhenc-Gelas M, Fiessinger JN. An abnormal antithrombin III (AT III) with low heparin affinity: AT III Clichy. Br J Haematol 1987; 66: 515-522
29 Koide T, Odani S, Takahashi K, Ono T, Sakuragawa N. Antithrombin III Toyama: replacement of arginine 47 by cysteine in hereditary abnormal antithrombin that lacks heparin-binding ability. Proc Natl Acad Sci, USA 1984; 81: 289-293
30 Brunei F, Duchange N, Fischer AM, Cohen GN, Zakin MM. Antithrombin III Alger: a new case of Arg 47 Cys mutation. Am J Haematol 1987; 25: 223-224
31 Borg JY, Owen MC, Soria C, Soria J, Caen J, Carrell RW. Proposed heparin binding site in antithrombin based on arginine 47. A new variant Rouen II, 47 Arg to Ser. J Clin Invest 1988 81. 1292-1296
32 Me Kay EG. A simple two-step procedure for the isolation of antithrombin III from biological fluid. Thromb Res 1981; 21: 375-382