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Thromb Haemost 1993; 70(05): 747-752
DOI: 10.1055/s-0038-1649663
Clinical Studies
Schattauer GmbH Stuttgart

Compound Heterozygosity in a Family with Protein C Deficiency Illustrating the Complexity of the Underlying Molecular Mechanism

Sophie Gandrille
1   Groupe de Recherche sur la Thrombose, INSERM CJF 91–01, UFR des Sciences Pharmaceutiques et Biologiques, Paris
,
Brigitte Jude
2   Laboratoire d'Hématologie et Service de Cardiologie C, Hôpital Cardiologique, Lille, France
,
Martine Alhenc-Gelas
1   Groupe de Recherche sur la Thrombose, INSERM CJF 91–01, UFR des Sciences Pharmaceutiques et Biologiques, Paris
,
Alain Millaire
2   Laboratoire d'Hématologie et Service de Cardiologie C, Hôpital Cardiologique, Lille, France
,
Martine Aiach
1   Groupe de Recherche sur la Thrombose, INSERM CJF 91–01, UFR des Sciences Pharmaceutiques et Biologiques, Paris
› Author Affiliations
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Publication History

Received 15 January 1993

Accepted after revision 09 June 1993

Publication Date:
05 July 2018 (online)

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Summary

The association of two missense mutations, a Leu 223 to Phe and an lie 403 to Met, is described in a family presenting with various protein C deficiency phenotypes. In this family, two subjects were compound heterozygotes with protein C levels of about 25%, the other members being heterozygous for only one of the mutations. The Leu 223 to Phe mutation was also found in 9 members of 3 other families and, in all cases but one, resulted in protein C levels below 60% associated with a high incidence of thrombotic complications. The other mutation, an lie 403 to Met, was identified in those of the family’ members who presented with borderline protein C concentrations. In such a family, the genomic DNA analysis represents the only way to differentiate between the genetic status of each family member. The results highlight the importance of the genotype determination and the poor discriminative power of the plasma assays currently used.

 

  • References

  • 1 Stenflo J. The biochemistry of protein C. In: Protein C and related proteins. Bertina RM. (ed). Churchill Livingstone: 1988: 21-54
    Google Scholar
  • 2 Foster DC, Yoshitake S, Davie EW. The nucleotide sequence of the gene for human protein C. Proc Natl Acad Sci USA 1985; 82: 4673-4677
    CrossrefPubMedGoogle Scholar
  • 3 Plutzky J, Hoskins JH, Long GL, Crabtree GR. Evolution and organization of the human protein C gene. Proc Natl Acad Sci USA 1986; 83: 546-550
    CrossrefPubMedGoogle Scholar
  • 4 Griffin JH, Evatt B, Zimmerman TS, Kleiss AJ, Wideman C. Deficiency of protein C in congenital thrombotic disease. J Clin Invest 1981; 68: 1370-1373
    CrossrefPubMedGoogle Scholar
  • 5 Broekmans AW, Veltkamp JJ, Bertina RM. Congenital protein C deficiency and venous thromboembolism. A study of three Dutch families. N Engl J Med 1983; 309: 340-344
    CrossrefPubMedGoogle Scholar
  • 6 Seligsohn U, Berger A, Abend M, Rubin L, Attias O, Zivelin A, Rapaport S. Homozygous protein C deficiency manifested by massive venous thrombosis in the newborn. N Engl J Med 1984; 310: 559-562
    CrossrefPubMedGoogle Scholar
  • 7 Bovill EG, Bauer KA, Dickerman JD, Callas P, West P. The clinical spectrum of heterozygous protein C deficiency in a large New England kindred. Blood 1989; 73: 712-717
    PubMedGoogle Scholar
  • 8 Allaart CF, Poort SR, Rosendaal FR, Reitsma PH, Bertina RM, Brict E. Increased risk of venous thrombosis in carriers of hereditary protein C deficiency defect. Lancet 1993; 341: 134-138
    CrossrefPubMedGoogle Scholar
  • 9 Marlar RA, Montgomery RR, Broekmans AW. and the Working party. Diagnosis and treatment of homozygous protein C deficiency. J Pediatr 1989; 114: 528-534
    CrossrefPubMedGoogle Scholar
  • 10 Marciniak E, Wilson HD, Marlar RA. Neonatal purpura fulminans: a genetic disorder related to the absence of protein C in blood. Blood 1985; 65: 15-20
    PubMedGoogle Scholar
  • 11 Tripodi A, Franchi F, Krachmalnicoff A, Mannucci PM. Asymptomatic homozygous protein C deficiency. Acta Haematol 1990; 83: 152-155
    CrossrefPubMedGoogle Scholar
  • 12 Tuddenham EGD, Takase T, Thomas AE, Awidi AS, Madanat FE, Abu HajirMM, Kernoff PBA, Hoffbrand AV. Homozygous protei C deficiency with delayed onset of symptoms at 7 to 10 months. Thromb Res 1989; 53: 475-484
    CrossrefPubMedGoogle Scholar
  • 13 Grundy CB, Lindo V, Kakkar W, Melissari E, Scully MF, Cooper DN. Late-onset homozygous protein C deficiency. Lancet 1991; 338: 575-576
    PubMedGoogle Scholar
  • 14 Conard J, Horellou MH, Van Dreden, Samama M, Reitsma PH, Poort S, Bertina RM. Homozygous protein C deficiency with late onset and recurrent coumarin-induced skin necrosis. Lancet 1992; 339: 743-744
    CrossrefPubMedGoogle Scholar
  • 15 Miletich J, Sherman L, Broze G. Absence of thrombosis in subjects with heterozygous protein C deficiency. N Engl J Med 1987; 317: 991-996
    CrossrefPubMedGoogle Scholar
  • 16 Reitsma PH, Poort SR, Bernardi F, Gandrille S, Long GL, Sala N, Cooper DN. Protein C deficiency: a database of mutations. Thromb Haemostas 1993; 69: 77-84
    PubMedGoogle Scholar
  • 17 Crabtree GR, Plutzky J, Marler R, Bertina RM, Broekmans AW, Griffin J, Zarcharski L, Gruppo R, Sala N, Long G. The range of genotypes underlying human protein C deficiency. Thromb Haemostas 1985; 54: 56 (abstract S331)
    PubMedGoogle Scholar
  • 18 Romeo G, Hassan HJ, Staempfli S, Roncuzzi L, Cianetti L, Leonardi A, Vicente V, Mannucci PM, Bertina RM, Peschle C, Cortese R. Hereditary thrombophilia: identification of nonsense and missense mutations in the protein C gene. Proc Natl Acad Sci USA 1987; 84: 2829-2832
    CrossrefPubMedGoogle Scholar
  • 19 Reitsma PH, Poort SR, Allaart CF, Briet E, Bertina RM. The spectrum of genetic defects in a panel of 40 Dutch families with symptomatic protein C deficiency type I: Heterogeneity and founder effects. Blood 1991; 78: 890-894
    PubMedGoogle Scholar
  • 20 Gandrille S, Vidaud M, Aiach M, Alhenc-Gelas M, Fischer AM, Gouault-Heilman M, Toulon P, Goossens M. Six previously undescribed mutations in 9 families with protein C quantitative deficiency. Thromb Haemost 1991; 65: 646 (abstract 3)
    PubMedGoogle Scholar
  • 21 Gandrille S, Vidaud M, Aiach M, Alhenc-Gelas M, Fischer AM, Gouault-Heilman M, Toulon P, Fiessinger JN, Goossens M. Two novel mutations responsible for hereditary type I protein C deficiency: Characterization by denaturing gradient gel electrophoresis. Human Mutation 1992; 1: 491-500
    CrossrefPubMedGoogle Scholar
  • 22 Grundy CB, Melissari E, Kakkar VV, Cooper DN. A molecular genetic study of protein C deficiency. Thromb Haemostas 1991; 65: 646 (abstract 2)
    PubMedGoogle Scholar
  • 23 Tsuda S, Reitsma PH, Miletich J. Molecular defect causing heterozygous protein C deficiency in three asymptomatic kindreds. Thromb Haemostas 1991; 65: 467 (abstract)7
    PubMedGoogle Scholar
  • 24 Yamamoto K, Tanimoto M, Matsushita T, Sigiura I, Takamatsu J, Saito H. Hereditary protein C deficiency due to a frameshift mutation in exon IX of the protein C gene. Thromb Haemostas 1991; 65: 646 (abstract 4)
    PubMedGoogle Scholar
  • 25 Sala N, Poort SR, Bertina RM, Soria JM, Fontcuberta J, Reitsma PH. Identification of two deletions and four point mutations in the protein C gene of 6 unrelated Spanish patients with hereditary protein C deficiency. Thromb Haemostas 1991; 65: 1197 (abstract 1809)
    PubMedGoogle Scholar
  • 26 Yamamoto K, Matsushita T, Sugiura I, Takamatsu J, Iwasaki E, Wada H, Deguchi K, Shirakawa S, Saito H. Homozygous protein C deficiency: identification of a novel missense mutation that causes impaired secretion of the mutant protein C. J Lab Clin Med 1992; 119: 682-689
    PubMedGoogle Scholar
  • 27 Grundy CB, Schulman S, Krawczak M, Kobosko J, Kakkar VV, Cooper DN. Protein C deficiency and thromboembolism: recurrent mutation at Arg 306 in the protein C gene. Hum Genet 1992; 88: 586-588
    CrossrefPubMedGoogle Scholar
  • 28 Sugahara Y, Miura O, Yuen P, Aoki N. Protein C deficiency Hong Kong 1 and 2: Hereditary protein C deficiency caused by two mutant alleles, a 5-nucleotide deletion and a missense mutation. Blood 1992; 80: 126-133
    PubMedGoogle Scholar
  • 29 Molho-Sabatier P, Aiach M, Gaillard I, Fiessinger JN, Fischer AM, Chadeuf G, Clauser E. Molecular characterization of antithrombin III (AT III) variants using polymerase chain reaction. Identification of the AT III Charleville as an Ala 384 Pro mutation. J Clin Invest 1989; 84: 1236-1242
    CrossrefPubMedGoogle Scholar
  • 30 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: 489-491
    PubMedGoogle Scholar
  • 31 Myers RM, Fischer SG, Lerman LS, Maniatis R. Nearly all single base substitutions in DNA fragments joined to a GC-clamp can be detected by denaturing gradient gel electrophoresis. Nucl Acids Res 1985; 13: 3131-3145
    CrossrefPubMedGoogle Scholar
  • 32 Myers RM, Fischer SG, Maniatis T, Lerman LS. Modification of the melting properties of duplex DNA by attachment of a G + C-rich DNA sequence as determined by denaturing gradient gel electrophoresis. Nucl Acids Res 1985; 13: 3111-3129
    CrossrefPubMedGoogle Scholar
  • 33 Sheffield VC, Cox DR, Lerman LS, Myers RM. Attachment of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. Proc Natl Acad Sci USA 1989; 86: 232-236
    CrossrefPubMedGoogle Scholar
  • 34 Lerman LS, Silverstein K. Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Methods in Enzymology 1987; 155: 482-501
    PubMedGoogle Scholar
  • 35 Attree O, Vidaud D, Vidaud M, Amselem S, Lavergne JM, Goossens M. Mutations in the catalytic domain of human coagulation factor IX: rapid characterization by direct genomic sequencing of DNA fragments displaying an altered melting behavior. Genomics 1989; 4: 266-272
    CrossrefPubMedGoogle Scholar
  • 36 Gyllensten UB, Erlich HA. Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sci USA 1988; 85: 7652-7656
    CrossrefPubMedGoogle Scholar
  • 37 Gandrille S, Aiach M. Polymorphism in the protein C gene detected by denaturing gradient gel electrophoresis. Nucl Acid Res 1991; 19: 6982
    PubMedGoogle Scholar
  • 38 Brantly M, Nukiwa T, Crystal RG. Molecular basis of alpha-1-antitrypsin deficiency. Amer J Med 1988; 84 (Suppl. 06) 13-31
    CrossrefPubMedGoogle Scholar
  • 39 Tait RC, Walker ID, Islam SI, Mitchell RM, Conkie JA, McCall F, Davidson F. Age related changes in protein C-activity in healthy adult males. Thromb Haemostas 1991; 65: 326-327
    PubMedGoogle Scholar
  • 40 Bottema CDK, Ketterling RP, Ii S, Yoon HS, Philips II I, Sommer SS. Missense mutations and evolutionary conservation of amino acids. Evidence that many of the amino acids in factor IX function as “spacer” elements. Amer J Hum Genet 1991; 49: 820-838
    PubMedGoogle Scholar
  • 41 Long GI, Belagaje RM, MacGillivray RTA. Cloning and sequencing of liver cDNA coding for bovine protein C. Proc Natl Acad Sci USA 1984; 81: 5653-5656
    CrossrefPubMedGoogle Scholar
  • 42 Tada N, Sato M, Tsujirama T, Iwase R, Hashimoto-Gotoh T. Isolation and characterization of a mouse protein C cDNA. J Biochem 1992; 111: 491-495
    CrossrefPubMedGoogle Scholar
  • 43 Giannelli F, Green PM, High KA, Sommer S, Lillicrap DP, Ludwig M, Olek K, Reitsma PH, Goossens M, Yoshioka A, Brownlee GG. Haemophilia B: database of point mutations and short additions and deletions Third edition. Nucl Acid Res 1992; 20 (suppl) 2027-2063
    CrossrefPubMedGoogle Scholar