Download PDF
Thromb Haemost 2000; 83(01): 86-92
DOI: 10.1055/s-0037-1613762
Commentary
Schattauer GmbH

Thirty-three Novel Mutations in the Protein C Gene

Martine Alhenc-Gelas
1   From the Laboratoire d’Hémostase, Hôpital Broussais - AP-HP and Unité INSERM 428, Faculté de Pharmacie, Université René Descartes, Paris, France
,
Sophie Gandrille
1   From the Laboratoire d’Hémostase, Hôpital Broussais - AP-HP and Unité INSERM 428, Faculté de Pharmacie, Université René Descartes, Paris, France
,
Marie-Laurence Aubry
1   From the Laboratoire d’Hémostase, Hôpital Broussais - AP-HP and Unité INSERM 428, Faculté de Pharmacie, Université René Descartes, Paris, France
,
Martine Aiach
1   From the Laboratoire d’Hémostase, Hôpital Broussais - AP-HP and Unité INSERM 428, Faculté de Pharmacie, Université René Descartes, Paris, France
,
deficiencies and the French INSERM network on molecular abnormalities responsible for protein C and protein S › Author Affiliations
Further Information

Publication History

Received 12 May 1999

Accepted after revision 02 August 1999

Publication Date:
06 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

We analyzed the protein C gene (PROC) with the denaturing gradient gel electrophoresis (DGGE) scanning strategy in a series of 129 patients with suspected protein C (PC) deficiency (93 with low plasma PC levels and 36 with borderline level). At least one sequence variation was found in 104 of the 129 patients. Thirty-nine sequence variations (found in 72 patients) were already reported detrimental mutations. Thirty-three were novel sequence variations, of which 19 (found in 25 patients) were probably detrimental. Five novel mutations (A1T, R9H, S11R, S12R and K193Q) were associated with qualitative plasma PC deficiency, suggesting or confirming the functional importance of amino acids at these positions. This strategy confirmed the diagnosis of inherited PC deficiency in 79/93 (84.9%) patients with low plasma PC levels and 14/36 (38.8%) patients with borderline values. In order to explain abnormal PC levels observed in patients who did not carry detrimental mutations, screening for the -1654C/T and -1641A/G PROC promoter polymorphisms known to influence plasma PC concentrations was performed. The frequency of the CG allele associated with lower PC concentrations was slightly but not significantly lower in 82 heterozygotes for detrimental PROC gene mutations than in 36 patients with no identified detrimental mutations.

Keywords

Protein C - thrombosis - coagulation - gene

* J.F. Abgrall (Hôpital Augustin Morvan, Brest, France), M.F. Aillaud and I. Juhan-Vague (CHU Timone, Marseille, France), N. Ajzenberg and M. Dreyfus (Hôpital Bicêtre, Le Kremlin-Bicêtre, France), M. Berruyer and M.C. Trzeciak (Hôpital Edouard Herriot, Lyon, France), C. Boinot (CHU La Miletrie, Poitiers, France), F. Bridey and D. de Prost (Hôpital Bichat, Paris, France), S. Brun (Hôpital Gaston Douvergne, Nîmes, France), M. David (Hôpital Ste-Justine, Montréal, Canada), E. de Maistre and T. Lecompte (CHU de Nancy, France), P. de Moerloose and G. Reber (Hôpital Cantonal Universitaire, Geneva, Switzerland), M.H. Denninger (Hôpital Beaujon, Clichy, France), E. Dupuy (Hôpital Lariboisière, Paris, France), J. Emmerich, J.N. Fiessinger, A. Jaque, A. Rance, M.E. Sirieix and J.F. Vitoux (Hôpital Broussais, Paris, France), M. Gouault-Heilmann (Hôpital Henri Mondor, Créteil, France), B. Jude (Hôpital Cardiologique, Lille, France), A. Le Querrec (CHU de la Côte de Nacre, Caen, France), M. Mexme (Orléans, France), A. Michaud-Mallet (Institut Arnault Tzanck, St-Laurent du Var, France), U. Nowak Gottl (Westfälische Wilhems Universität, Münster, Germany), P. Priollet (Hôpital St-Joseph, Paris, France), J. Reynaud (Hôpital Nord, Saint-Etienne, France), V. Siguret (Hôpital Charles Foix, Ivry, France), B. Tardy (Hôpital Bellevue, Saint-Etienne, France), P. Toulon (Hôpital Cochin, Paris, France), N.H. Toumi (Hôpital d’Enfants, Tunis, Tunisie), F. Truchaud (Institut de Biologie des Hôpitaux, Nantes, France), C. Vergnes (Hôpital Cardiologique du Haut-Lévêque, Pessac, France), M. Wolf (Hôpital Antoine Béclère, Clamart, France).


 

  • References

  • 1 Esmon TS. Protein C: Biochemistry, physiology, and clinical implications. Blood 1983; 62: 1155-8.
    PubMedSearch in Google Scholar
  • 2 Walker FG. Regulation of activated protein C by protein S. The role of phospholipids in factor Va inactivation. J Biol Chem 1981; 256: 11128-31.
    PubMedSearch in Google Scholar
  • 3 Kato A, Miura O, Sumi Y, Aoki N. Assignment of the protein C gene (PROC) to chromosome 2q14:q21. Cytogenet Cell Genet 1988; 47: 46-7.
    CrossrefPubMedSearch in Google Scholar
  • 4 Patracchini P, Aiello V, Palazzi P, Calzolari E, Bernardi F. Sublocalization of the human protein C gene on chromosome 2q13-q14. Hum Genet 1989; 81: 191-2.
    CrossrefPubMedSearch in Google Scholar
  • 5 Foster DC, Yoshitake S, Davie EW. The nucleotide sequence of the gene for human protein C. Proc Natl Acad Sci USA 1985; 82: 4673-7.
    CrossrefPubMedSearch in Google Scholar
  • 6 Plutsky J, Hoskins JA, Long GL, Crabtree GR. Evolution and organization of the human protein C gene. Proc Natl Acad Sci USA 1986; 83: 546-50.
    CrossrefPubMedSearch in Google Scholar
  • 7 Stenflo J. The biochemistry of protein C. in Bertina RM. (ed): Protein C and related proteins. Edinburgh, London, Melbourne, New York: Churchill Livingstone; 1988: 21-41.
    Search in Google Scholar
  • 8 Spek CA, Poort SR, Bertina RM, Reitsma PH. Determination of the allelic and haplotype frequencies of three polymorphisms in the promoter region of the human protein C gene. Blood Coag Fibrinol 1994; 05: 309-11.
    CrossrefPubMedSearch in Google Scholar
  • 9 Spek CA, Koster T, Rosendaal FR, Bertina RM, Reitsma PH. Genotypic variation in the promoter region of the protein C gene is associated with plasma protein C levels and thrombotic risk. Arterioscler Thromb Vasc Biol 1995; 15: 214-8.
    CrossrefPubMedSearch in Google Scholar
  • 10 Aiach M, Nicaud V, Alhenc-Gelas M, Gandrille S, Arnaud E, Amiral J, Guize L, Fiessinger JN, Emmerich J. Complex association of protein C gene promoter polymorphism with circulating protein C levels and the thrombotic risk. Arterioscler Thromb Vasc Biol 1999; 19: 1573-6.
    CrossrefPubMedSearch in Google Scholar
  • 11 Reitsma PH, Bernardi F, Doig RG, Gandrille S, Greengard JS, Ireland H, Krawczak M, Lind B, Long GL, Poort SR, Saito H, Sala N, Witt I, Cooper DN. Protein C deficiency: a database of mutations, 1995 Update. Thromb Haemost 1995; 73: 876-89.
    Thieme ConnectPubMedSearch in Google Scholar
  • 12 Allaart CF, Poort SR, Rosendaal FR, Reitsma PH, Bertina RM, Briet E. Increased risk of venous thrombosis in carriers of hereditary protein C deficiency defect. Lancet 1993; 341: 134-8.
    CrossrefPubMedSearch in Google Scholar
  • 13 Miletich J, Sherman L, Broze GJ. Absence of thrombosis in subjects with heterozygous protein C deficiency. N Engl J Med 1987; 317: 991-6.
    CrossrefPubMedSearch in Google Scholar
  • 14 Tait RC, Walker ID, Reitsma PH, Islam SIAM, McCall F, Poort SR, Conkie JA, Bertina RM. Prevalence of protein C deficiency in the healthy population. Thromb Haemost 1995; 73: 87-92.
    Thieme ConnectPubMedSearch in Google Scholar
  • 15 Gladson CL, Scharrer I, Hach V, Beck KH, Griffin JH. The frequency of type I heterozygous protein S and protein C deficiency in 141 unrelated young patients with venous thrombosis. Thromb Haemost 1988; 59: 18-22.
    Thieme ConnectPubMedSearch in Google Scholar
  • 16 Broekmans AW, Van der Lincen IK, Veltkamp JJ, Bertina TM. Prevalence of isolated protein C deficiency in patients with venous thromboembolic disease and in the population. Thromb Haemost 1983; 50: 350a.
    PubMedSearch in Google Scholar
  • 17 Marlar RA, Montgomery RR, Broekmans AW. Report on the diagnosis and treatment of homozygous protein C deficiency. Report of the working party on homozygous protein C deficiency of the ICTH-Subcommittee on Protein C and Protein S. Thromb Haemost 1989; 61: 529-31.
    Thieme ConnectPubMedSearch in Google Scholar
  • 18 Gandrille S, Aiach M. and the French INSERM network on Molecular abnormalities responsible for protein C and protein S deficiencies. Identification of mutations in 90 of 121 consecutive symptomatic French patients with a type I protein C deficiency. Blood 1995; 86: 2598-605.
    PubMedSearch in Google Scholar
  • 19 Gandrille S, Goossens M, Aiach M. Scanning method to establish the molecular basis of protein C deficiencies. Hum Mutat 1994; 04: 20-30.
    CrossrefPubMedSearch in Google Scholar
  • 20 Gandrille S, Alhenc-Gelas M, Aiach M. A rapid screening method for the factor V Arg 506 Gln mutation. Blood Coag Fibrinol 1995; 06: 245-9.
    CrossrefPubMedSearch in Google Scholar
  • 21 Alhenc-Gelas M, Arnaud E, Nicaud V, Aubry ML, Fiessinger JN, Aiach M, Emmerich J. Venous thromboembolic disease and the prothrombin, methylene tetrahydrofolate reductase and factor V genes. Thromb Haemost 1999; 81: 506-10.
    Thieme ConnectPubMedSearch in Google Scholar
  • 22 Okafuji T, Maekawa K, Nawa K, Marumoto Y. The cDNA cloning and mRNA expression of rat protein C. Biochim Biophys Acta 1992; 1131: 329-32.
    CrossrefPubMedSearch in Google Scholar
  • 23 Tada N, Sato M, Tsujimura A, Iwase R, Hashimoto-Gotoh T. Isolation and characterization of a mouse protein C cDNA. J Biochem 1992; 111: 491-5.
    CrossrefPubMedSearch in Google Scholar
  • 24 Long GL, Belagaje RM, Mac RTAGillivray. Cloning and sequencing of liver cDNA coding for bovine protein C. Proc Natl Acad Sci USA 1984; 81: 5653-6.
    CrossrefPubMedSearch in Google Scholar
  • 25 Bancroft JD, McDowell SA, Friezner-Degen SJ. The human prothrombin gene: transcriptional regulation in HepG2 cells. Biochemistry 1992; 31: 12469-76.
    CrossrefPubMedSearch in Google Scholar
  • 26 O’Hara PJ, Grant FJ, Haldeman BA, Gray CL, Insley MY, Hagen FS, Murray MJ. Nucleotide sequence of the gene coding for human factor VII, a vitamin K-dependent protein participating in blood coagulation. Proc Natl Acad Sci USA 1987; 84: 5158-62.
    CrossrefPubMedSearch in Google Scholar
  • 27 Yoshitake S, Schach BG, Foster DC, Davie EW, Kurachi K. Nucleotide sequence of the gene for human factor IX (antihemophilic factor B). Biochemistry 1985; 24: 3736-50.
    CrossrefPubMedSearch in Google Scholar
  • 28 Leytus SP, Foster DC, Kurachi K, Davie EW. Gene for human factor X: a blood coagulation factor whose organization is essentially identical with that of factor IX and protein C. Biochemistry 1986; 25: 5098-102.
    CrossrefPubMedSearch in Google Scholar
  • 29 Schmidel DK, Tatro AV, Phelps AG, Tomczak JA, Long GL. Organization of the human protein S genes. Biochemistry 1990; 29: 7845-52.
    CrossrefPubMedSearch in Google Scholar
  • 30 Foster DC, Rudinski MS, Schach BG, Berkner KL, Kumar AA, Hagen FS, Sprecher CA, Insley MY, Davie EW. Propeptide of human protein C is necessary for γ-carboxylation. Biochemistry 1987; 26: 7003-11.
    CrossrefPubMedSearch in Google Scholar
  • 31 Mullien P, Balland A, Lepage P, Mischler F, Dott K, Hauss C, Grandgeorge M, Lecocq JP. Increased biological activity of a recombinant factor IX variant carrying alanine at position +1. Protein Eng 1990; 03: 629-33.
    CrossrefPubMedSearch in Google Scholar
  • 32 Geng JP, Castellino FJ. The propeptides of human protein C, factor VII, and factor IX are exchangeable with regard to directing gamma-carboxylation of these proteins. Thromb Haemost 1996; 76: 205-7.
    Thieme ConnectPubMedSearch in Google Scholar
  • 33 Zhang L, Castellino FJ. The contributions of individual γ-carboxyglutamic acid residues in the calcium-dependent binding of recombinant human protein C to acidic phospholipid vesicles. J Biol Chem 1993; 268: 12040-5.
    PubMedSearch in Google Scholar
  • 34 Shen L, Shah AM, Dahlback B, Nelsestuen GL. Enhancement of human protein C function by site directed mutagenesis of the gamma-carboxyglutamic acid domain. J Biol Chem 1998; 273: 31086-91.
    CrossrefPubMedSearch in Google Scholar
  • 35 Gerlitz B, Grinnell BW. Mutation of protease domain residues Lys 37-39 in human protein C inhibits activation by the thrombomodulin-thrombin complex without affecting activation by free thrombin. J Biol Chem 1996; 271: 22285-8.
    CrossrefPubMedSearch in Google Scholar
  • 36 Tsay W, Lee YM, Lee SC, Shen MC, Chen PJ. Characterization of human protein C gene promoter: insights from natural human mutants. DNA and Cell Biol 1996; 15: 907-19.
    CrossrefPubMedSearch in Google Scholar
  • 37 Cooper DN, Youssoufian H. The CpG dinucleotide and human genetic disease. Hum Genet 1998; 78: 151-5.
    PubMedSearch in Google Scholar
  • 38 Ireland H, Thompson E, Lane DA. and the protein C study group. Gene mutations in 21 unrelated cases of phenotypic heterozygous protein C deficiency and thrombosis. Thromb Haemost 1996; 76: 867-73.
    Thieme ConnectPubMedSearch in Google Scholar
  • 39 Gejman PV, Cao Q, Guedj F, Sommer S. The sensitivity of denaturing gradient gel electrophoresis: a blinded analysis. Mutat Res 1998; 382: 109-14.
    PubMedSearch in Google Scholar
  • 40 Macek Jr M, Mercier B, Maekova A, Miller PW, Hamosh A, Ferec C, Cutting GR. Sensitivity of the denaturing gradient gene electrophoresis technique in detection of known mutations and novel Asian mutations in the CFTR gene. Hum Mutat 1997; 09: 136-47.
    CrossrefPubMedSearch in Google Scholar
  • 41 Alhenc-Gelas M, Emmerich J, Gandrille S, Aubry ML, Benaily N, Fiessinger JN, Aiach M. Protein C infusion in a patient with inherited protein C deficiency caused by two missense mutations: Arg 178 to Gln and Arg-1 to His. Blood Coag Fibrinol 1995; 06: 35-42.
    CrossrefPubMedSearch in Google Scholar
  • 42 Monagle P, Andrew M, Halton J, Marlar R, Jardine L, Vegh P, Johnston M, Webber C, Masicotte MP. Homozygous protein C deficiency: description of a new mutation and successful treatment with low molecular weight heparin. Thromb Haemost 1998; 79: 756-61.
    Thieme ConnectPubMedSearch in Google Scholar
  • 43 Kemahli S, Alhenc-Gelas M, Gandrille S, Aiach M, Akar N, Cin S. Homozygous protein C deficiency with a double variant His 202 to Tyr and Ala 346 to Thr. Blood Coag Fibrinol 1998; 09: 351-4.
    CrossrefPubMedSearch in Google Scholar