Download PDF
Thromb Haemost 1998; 79(02): 249-253
DOI: 10.1055/s-0037-1614973
Letters to the Editor
Schattauer GmbH

Antithrombin Cambridge II (Ala384Ser): Clinical, Functional and Haplotype Analysis of 18 Families

D. J. Perry
1   From the Haemophilia Centre and Haemostasis Unit, Department of Haematology, the Royal Free Hospital and School of Medicine, London
,
M. E. Daly
2   From the Department of Medicine and Pharmacology, Royal Hallamshire Hospital, Sheffield
,
R. C. Tait
3   From the Department of Haematology, Glasgow Royal Infirmary, Glasgow
,
I. D. Walker
3   From the Department of Haematology, Glasgow Royal Infirmary, Glasgow
,
K. Brown
4   From the Department of Haematology, University of Cambridge, MRC Centre, Cambridge
,
N. J. Beauchamp
2   From the Department of Medicine and Pharmacology, Royal Hallamshire Hospital, Sheffield
,
F. E. Preston
2   From the Department of Medicine and Pharmacology, Royal Hallamshire Hospital, Sheffield
,
H. Gyde
5   From the Department of Haematology, Birmingham Heartlands Hospital, Birmingham, UK
,
P. L. Harper
4   From the Department of Haematology, University of Cambridge, MRC Centre, Cambridge
,
R. W. Carrell
4   From the Department of Haematology, University of Cambridge, MRC Centre, Cambridge
› Author AffiliationsThe financial support of the Wellcome Trust, The British Heart Foundation and Medical Research Council is gratefully acknowledged. DJP is supported by the Katharine Dormandy Trust for Haemophilia and Allied Disorders.
Further Information

Publication History

Received 27 June 1996

Accepted after resubmission 16 September 1997

Publication Date:
08 December 2017 (online)

    Permissions and Reprints

Summary

Thirty-one individuals from 18 unrelated families with antithrombin deficiency have been identified as having a single point mutation within codon 384 (13268 GCA→TCA) resulting in an alanine to serine substitution. Six families (11 individuals) were identified by the screening of individuals with thromboembolic disease or with a family history of thromboembolic disease, whilst the remaining 12 families (20 individuals) were identified by screening of asymptomatic blood donors. Four individuals had a history of venous thrombotic disease, a further 2 gave a history of superficial thrombophlebitis but the remaining 25 individuals were asymptomatic. Affected individuals demonstrated normal immunological levels of antithrombin but a decrease in anti-IIa activity in the presence of heparin. Haplotype analysis was used to examine the possibility of a founder effect to explain the high frequency of this non-CpG mutation. 29/31 individuals showed a single common “core” haplotype, the only variation existing in the number of copies of an (ATT)n repeat polymorphism – 13, 14, 15 or 17. The results suggest that at most there are four independent origins for this mutation.

 

  • References

  • 1 Rosenberg RD, Damus PS. The purification and mechanism of action of human antithrombin-heparin cofactor. J Biol Chem 1973; 248: 6490-505.
    PubMedGoogle Scholar
  • 2 Damus PS, Hicks M, Rosenberg RD. Anticoagulant action of heparin. Nature 1973; 246: 355-7.
    CrossrefPubMedGoogle Scholar
  • 3 Kurachi K, Fujikawa K, Schmer G, Davie EW. Inhibition of bovine factor IXa and factor X by antithrombin III. Biochemistry 1976; 15: 373-7.
    CrossrefPubMedGoogle Scholar
  • 4 Abildgaard U. Binding of thrombin to antithrombin III. Scand J Clin Lab Invest 1969; 24: 23-7.
    CrossrefPubMedGoogle Scholar
  • 5 Frebelius S, Nydahl S, Swedenborg J. Coagulant and noncoagulant thrombin enzymatic activity on the endothelium. Blood Coagul Fibrinolysis 1990; 1 (03) 285-92.
    CrossrefPubMedGoogle Scholar
  • 6 Bock SC, Harris JF, Balazs I, Trent JM. Assignment of the human antithrombin III structural gene to chromosome 1q23-25. Cytogenet Cell Genet 1985; 39 (01) 67-9.
    CrossrefPubMedGoogle Scholar
  • 7 Olds RJ, Lane DA, Chowdhury V, DeStefano V, Leone G, Thein S-L. Complete nucleotide sequence of the antithrombin gene. Evidence for homologous recombination causing thrombophilia. Biochemistry 1993; 32: 4216-24.
    CrossrefPubMedGoogle Scholar
  • 8 Hunt LT, Dayhoff MO. A surprising new protein superfamily containing ovalbumin, antithrombin-III and alpha 1-proteinase inhibitor. Biochem Biophys Res Commun 1980; 95: 864-71.
    CrossrefPubMedGoogle Scholar
  • 9 Carrell RW, Boswell DR. Serpins: the superfamily of plasma serine proteinase inhibitors. In: Proteinase Inhibitors. Barrett A, Salvesen G. eds. Amsterdam: Elsevier Biomedical Press; 1986: 405-19.
    Google Scholar
  • 10 Harper PL, Luddington RJ, Daly M, Bruce D, Williamson D, Edgar PF, Perry DJ, Carrell RW. The incidence of dysfunctional antithrombin variants: four cases in 210 patients with thromboembolic disease. Br J Haematol 1991; 77 (03) 360-4.
    CrossrefPubMedGoogle Scholar
  • 11 Tait RC, Walker ID, Perry DJ, Islam SIAM, Daly ME, McCall F, Conkie JA, Carrell RW. Prevalence of antithrombin III deficiency in the healthy population. Br J Haematol 1994; 87: 106-12.
    CrossrefPubMedGoogle Scholar
  • 12 Thaler E, Lechner K. Antithrombin III deficiency and thromboembolism. Clinics in Haematology 1981; 10: 369-90.
    PubMedGoogle Scholar
  • 13 Lane DA, Olds RJ, Boisclair V, Chowdhury V, Thein SL, Cooper DN, Blajchman M, Perry DJ, Emmerich J, Aiach M. Antithrombin III Mutation Database: First Update. Thromb Haemost 1993; 70 (02) 361-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Perry DJ, Daly M, Harper PL, Tait RC, Price J, Walker ID, Carrell RW. Antithrombin Cambridge II 384 Ala to Ser. Further evidence of the role of the reactive centre loop in the inhibitory function of the serpins. FEBS Letts 1991; 285 (02) 248-50.
    CrossrefPubMedGoogle Scholar
  • 15 Laurell CB. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52.
    CrossrefPubMedGoogle Scholar
  • 16 Abildgaard U, Lie M, Ødegård OR. Antithrombin (Heparin Cofactor) Assay with “New” Chromogenic Substrates (S-2238 and Chromozym TH). Thromb Res 1977; 11: 549-53.
    CrossrefPubMedGoogle Scholar
  • 17 Ødegård OR, Lie M, Abildgaard U. Antifactor Xa activity measured with amidolytic methods. Haemostasis 1976; 5: 265-76.
    PubMedGoogle Scholar
  • 18 Harper PL, Daly M, Price J, Edgar PF, Carrell RW. Screening for heparin binding variants of antithrombin. Br J Haematol 1991; 44 (06) 477-9.
    PubMedGoogle Scholar
  • 19 Sas G, Pepper DS, Cash JD. Plasma and Serum Antithrombin-III: Differentiation by Crossed Immunoelectrophoresis. Thromb Res 1975; 6: 87-91.
    CrossrefPubMedGoogle Scholar
  • 20 Orita M, Suzuki Y, Sekiya T, Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using polymerase chain reaction. Genomics 1989; 5: 874-9.
    CrossrefPubMedGoogle Scholar
  • 21 Perry DJ, Carrell RW. Hydrolink Gels: a rapid and simple approach to the detection of DNA mutations in thromboembolic disease. J Clin Pathol 1992; 45: 158-60.
    CrossrefPubMedGoogle Scholar
  • 22 Bock SC, Levitan DJ. Characterization of an unusual DNA length polymorphism 5’ to the human antithrombin III gene. Nucl Acids Res 1983; 11 (24) 8569-82.
    CrossrefPubMedGoogle Scholar
  • 23 Bock SC, Radziejewska E. A NheI RFLP in the human antithrombin III gene (1q23-q25) (AT3). Nucl Acids Res 1991; 19: 2519.
    PubMedGoogle Scholar
  • 24 Prochownik EV, Markham AF, Orkin SH. Isolation of a cDNA clone for human antithrombin III. J Biol Chem 1983; 258 (13) 8389-94.
    PubMedGoogle Scholar
  • 25 Olds RJ, Lane DA, Chowdhury V, Sas G, Pabinger I, Meitinger T, Thein SL. (ATT) trinucleotide repeats in the antithrombin gene and their use in determining the origin of repeated mutations. Hum Mutat 1994; 4: 31-41.
    CrossrefPubMedGoogle Scholar
  • 26 Perry DJ. Trinucleotide repeat polymorphism in the human antithrombin III (AT3) gene. Hum Mol Genet 1993; 2 (05) 618.
    PubMedGoogle Scholar
  • 27 Daly ME, Perry DJ. DdeI polymorphism in intron 5 of the ATIII gene. Nucl Acids Res 1990; 18 (18) 5583.
    PubMedGoogle Scholar
  • 28 Wu S, Seino S, Bell GI. Human antithrombin III (AT3) gene length polymorphism revealed by the polymerase chain reaction. Nucl Acids Res 1989; 17 (15) 6433.
    CrossrefPubMedGoogle Scholar
  • 29 Newton GR, Graham A, Hepinstall LE, Powell SJ, Summers C, Kalsheker N, Smith JC, Markham AF. Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucl Acids Res 1989; 17: 2503-16.
    CrossrefPubMedGoogle Scholar
  • 30 Huber R, Carrell RW. Implications of the Three-Dimensional Structure of 1-Antitrypsin for Structure and Function of Serpins. Biochemistry 1989; 28 (23) 8951-65.
    CrossrefPubMedGoogle Scholar
  • 31 Carrell RW, Evans DLI. Serpins: mobile conformations in a family of proteinase inhibitors. Current Opinion in Structural Biology 1992; 2: 438-46.
    CrossrefPubMedGoogle Scholar
  • 32 Carrell RW, Stein PE, Fermi G, Wardell MR. Biological implications of a 3Å structure of dimeric antithrombin. Structure 1994; 2: 257-70.
    CrossrefPubMedGoogle Scholar
  • 33 Molho-Sabatier P, Aiach M, Gaillard I, Fiessinger JN, Fischer AM, Chadeuf G, Clauser E. Molecular Characterization of Antithrombin III (ATIII) Variants Using Polymerase Chain Reaction. J Clin Invest 1989; 84: 1236-42.
    CrossrefPubMedGoogle Scholar
  • 34 Perry DJ, Harper PL, Fairham S, Daly ME, Carrell RW. Antithrombin Cambridge, 384 Ala to Pro: A new variant idientified using the polymerase chain reaction. FEBS Letts 1989; 254 (1, 2) 174-6.
    CrossrefPubMedGoogle Scholar
  • 35 Devraj-Kizuk R, Chui DH, Prochownik EV, Carter CJ, Ofosu FA, Blajchman MA. Antithrombin-III-Hamilton: a gene with a point mutation (guanine to adenine) in codon 382 causing impaired serine protease reactivity. Blood 1988; 72 (05) 1518-23.
    PubMedGoogle Scholar
  • 36 Demers C, Henderson P, Blajchman MA, Wells MJ, Mitchell L, Johnston M, Ofosu FA, Fernandez RF, Andrew M, Hirsh J. An antithrombin III assay based on factor Xa inhibition provides a more reliable test to identify congenital antithrombin III deficiency than an assay based on thrombin inhibition. Thromb Haemostas 1993; 69 (03) 231-5.
    PubMedGoogle Scholar
  • 37 Ni H, Waye JS, Sheffield WP, Eng B, Blajchman MA. Genetic Linkage Studies in Antithrombin-Deficient Kindreds using a Highly Polymorphic Trinucleotide Short Tandem Repeat (STR) Within the Human Antithrombin Gene. Am J Hematol 1994; 46: 107-11.
    CrossrefPubMedGoogle Scholar
  • 38 Cooper DN, Krawczak M. The mutational spectrum of single base-pair substitutions causing human genetic disease: patterns and predictions. Hum Genet 1990; 85 (01) 55-74.
    PubMedGoogle Scholar