Biochemical activity and gene analysis of inherited protein C and antithrombin deficiency in two Chinese pedigrees

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Journal Title:
Volume 117, Issue 06, 2004
Key Word:
protein C;antithrombin;gene mutation;thrombosis

Abstract: Background We identified the gene mutations in two Chinese pedigree of type Ⅰ hereditary protein C deficiency and type Ⅰ hereditary antithrombin deficiency.Methods The plasma level of protein C activity (PC∶ A), protein C antigen (PC∶ Ag) , protein S activity, antithrombin activity (AT∶ A) and antithrombin antigen (AT∶ Ag) of propositi and two family members were detected using ELISA and chromogenic assay, respectively. All exons and intron-exon boundaries of protein C gene and antithrombin gene were analyzed by direct sequencing of the corresponding amplified PCR products in DNA from the propositus. Results The plasma PC∶ A and PC∶ Ag of propositus 1 was 26% and 1.43 mg/dl, respectively. The PC∶ Ag and PC∶ A of his father were normal. The decreased PC∶ A level was seen in his mother and 4 of his maternal pedigree. PS∶ A and AT∶ A were all normal in pedigree 1 members. A C5498T heterozygous mutation in exon 3 of protein C gene, resulting in the substitution of Arg for Trp at the 15th amino acid, was identified in propositus 1 and 8 of his relatives. The plasma AT∶ A and AT∶ Ag of propositus 2 was 48.6% and 10.4 mg/dl, respectively. The reduced AT∶ A and AT∶ Ag levels were found in his father and 5 of paternal pedigree. PC∶ A, PC∶ Ag and PS∶ A were all in normal range. A heterozygous 13387-9G deletion in exon 6 of antithrombin gene was identified in propositus 2. This mutation introduced a frameshift and a premature stop at codon 426 and existed in 6 members of pedigree 2.Conclusion The C5498T heterozygous mutation in exon 3 of protein C gene, first reported in China, leads to type I hereditary protein C deficiency. The 13387-9G deletion, a novel mutation, can cause antithrombin deficiency and thrombosis.

  • [1]Crowther MA, Kelton JG. Congenital thrombophilic states associated with venous thrombosis: A qualitative overview and proposed classification system. Ann Intern Med 2003;138:128-134.
  • [2]Martinelli I, Mannucci PM, De Stefano V, et al. Different risks of thrombosis in four coagulation defects associated with inherited thrombophilia: a study of 150 families. Blood 1998;92:2353-2358.
  • [3]Lind B, van Solinge WW, Schwartz M, et al. Splice site mutation in the human protein C gene associated with venous thrombosis: demonstration of exon skipping by ectopic transcript analysis. Blood 1993;82:2423-2432.
  • [4]Olds RJ, Lane DA, Chowhury V. et al. Complete nucleotide sequence of the antithrombin gene: evidence for homologous recombination causing thrombophilia. Biochemistry 1993;32:4216-4224.
  • [5]Reitsma PH, Bernardi F, Doig RG, et al. Protein C deficiency. A database of mutations, 1995 update. Thromb Haemost 1995;73:876-889.
  • [6]Zhang L, Jhingan A, Castellino FJ. Role of individual γ-carboxyglutamic acid residues of activated human protein C in defining its in vitro anticoagulant activity. Blood 1992;80:942-952.
  • [7]Tokunaga F, Tsukamoto T, Koide T. Cellular basis foe protein C deficiency caused by a single amino substitution at Arg 15 in the γ-carboxyglutamic acid domain. J Biochem 1996;120:360-368.
  • [8]Fitches AC, Lewandowski K, Olds R J. Creation of an additional glycosylation site as a mechanism for type 1 antithrombin deficiency. Thromb Haemost 2001;86:1023-1027.
  • [9]Bovil EG, Hasstedt SJ, Leppert MF, et al. Hereditary thrombophilia as a model for multigenic disease. Thromb Haemost 1999;82:662-666.
  • [10]Bauer KA, Rosendaal FR, Heit JA. Hypercoagulability: too many tests, too much conflicting data. Hematology 2002;353-368.
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