Download PDF
Thromb Haemost 2002; 88(01): 144-148
DOI: 10.1055/s-0037-1613167
Review Article
Schattauer GmbH

A Single Thymine Nucleotide Deletion Responsible for Congenital Deficiency of Plasmin Inhibitor

Haruhiko Yoshinaga
1   Department of Hematology
,
Masako Nakahara
2   Graduate School of Allied Health Sciences, Tokyo Medical and Dental University, Tokyo, Japan
,
Takatoshi Koyama
1   Department of Hematology
2   Graduate School of Allied Health Sciences, Tokyo Medical and Dental University, Tokyo, Japan
,
Aya Shibamiya
2   Graduate School of Allied Health Sciences, Tokyo Medical and Dental University, Tokyo, Japan
,
Fumie Nakazawa
2   Graduate School of Allied Health Sciences, Tokyo Medical and Dental University, Tokyo, Japan
,
Lindsey A. Miles
3   Department of Cell Biology, The Scripps Research Institute, La Jolla, CA, USA
,
Shinsaku Hirosawa
1   Department of Hematology
,
Nobuo Aoki
1   Department of Hematology
› Author Affiliations
Further Information

Publication History

Received 20 December 2001

Accepted after resubmission 18 April 2002

Publication Date:
09 December 2017 (online)

    Permissions and Reprints

Summary

Plasma plasmin inhibitor (PI) is a physiological inhibitor of plasminmediated fibrinolysis and constitutes a hemostatic component in blood plasma; hence its deficiency results in a severe hemorrhagic diathesis. We have carried out molecular analysis of American family members with congenital PI deficiency, and detected a single thymine deletion at nucleotide position 332 in exon 5. The deletion was found in both alleles of the homozygotes and in one allele of the heterozygotes, and the patterns of restriction fragment length polymorphism created by the mutation in the family members were compatible with their phenotypes. The deletion caused a frameshift leading to an alteration and shortening of the deduced amino acid sequence. The amino acid sequence consists of the first 83 amino acids of the N-terminal sequence of the normal PI and additional new amino acids, resulting in a mutant composed of 94 amino acids in contrast to 464 amino acids of the normal PI. In transient expression analysis, the mutant PI whose molecular size was compatible with the predicted amino acid sequence was detected in the lysates of the cells transfected with the mutated PI expression vector. The mutant PI was retained and underwent progressive degradation within the cells, and was minimally excreted into the media. These data indicate that this mutation is the cause of PI deficiency in this pedigree.

Keywords

Plasmin inhibitor (PI) - mutation - fibrinolysis - thymine deletion
 

  • References

  • 1 Blombäck M, Abildgaard U, van den Besselaar AM, Clementson KJ, Dahlbäck B, Exner T, Francis CW, Gaffney P, Gralnick H, Hoyer LW. et al. Nomenclature of quantities and units in thrombosis and haemostasis (recommendation 1993). A Collaborative project of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis (ISTH/SSC) and the Commission/Committee on Quantities and Units (in Clinical Chemistry) of the International Union of Pure and Applied Chemistry-International Federation of Clinical Chemistry (IUPAC-IFCC/CQU(CC)). Thromb Haemost 1994; 71: 375-94.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Moroi M, Aoki N. Isolation and characterization of α2-plasmin inhibitor from human plasma. A novel proteinase inhibitor which inhibits activatorinduced clot lysis. J Biol Chem 1976; 251: 5956-65.
    PubMedGoogle Scholar
  • 3 Wiman B, Collen D. Purification and characterization of human antiplasmin, the fast-acting plasmin inhibitor in plasma. Eur J Biochem 1977; 78: 19-26.
    CrossrefPubMedGoogle Scholar
  • 4 Bangert K, Johnsen AH, Christensen U, Thorsen S. Different N-terminal forms of α2-plasmin inhibitor in human plasma. Biochem J 1993; 291: 623-5.
    CrossrefPubMedGoogle Scholar
  • 5 Koyama T, Koike Y, Toyota S, Miyagi F, Suzuki N, Aoki N. Different NH2-terminal form with 12 additional residues of α2-plasmin inhibitor from human plasma and culture media of Hep G2 cells. Biochem Biophys Res Commun 1994; 200: 417-22.
    CrossrefPubMedGoogle Scholar
  • 6 Hirosawa S, Nakamura Y, Miura O, Sumi Y, Aoki N. Organization of the human α2-plasmin inhibitor gene. Proc Natl Acad Sci USA 1988; 85: 6836-40.
    CrossrefPubMedGoogle Scholar
  • 7 Kato A, Hirosawa S, Toyota S, Nakamura Y, Nishi H, Kimura A, Sasazuki T, Aoki N. Localization of the human α2-plasmin inhibitor gene (PLI) to 17p13. Cytogenet Cell Genet 1993; 62: 190-1.
    CrossrefPubMedGoogle Scholar
  • 8 Welch SK, Francke U. Assignment of the human α2-plasmin inhibitor gene (PLI) to chromosome 17, region pter-p12, by PCR analysis of somatic cell hybrids. Genomics 1992; 13: 213-4.
    CrossrefPubMedGoogle Scholar
  • 9 Aoki N, Moroi M, Matsuda M, Tachiya K. The behavior of α2-plasmin inhibitor in fibrinolytic states. J Clin Invest 1977; 60: 361-9.
    CrossrefPubMedGoogle Scholar
  • 10 Aoki N, Moroi M, Tachiya K. Effects of α2-plasmin inhibitor on fibrin clot lysis. Its comparison with α2-macroglobulin. Thromb Haemost 1978; 39: 22-31.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 11 Clemmensen I, Thorsen S, Mullertz S, Petersen LC. Properties of three different molecular forms of the α2-plasmin inhibitor. Eur J Biochem 1981; 120: 105-12.
    CrossrefPubMedGoogle Scholar
  • 12 Aoki N, Sakata Y, Ichinose A. Fibrin-associated plasminogen activation in α2-plasmin inhibitor deficiency. Blood 1983; 62: 1118-22.
    PubMedGoogle Scholar
  • 13 Sakata Y, Aoki N. Cross-linking of cx2-plasmin inhibitor to fibrin by fibrin-stabilizing factor. J Clin Invest 1980; 65: 290-7.
    CrossrefPubMedGoogle Scholar
  • 14 Tamaki T, Aoki N. Cross-linking of α2-plasmin inhibitor to fibrin catalyzed by activated fibrin-stabilizing factor. J Biol Chem 1982; 257: 14767-72.
    PubMedGoogle Scholar
  • 15 Kimura S, Aoki N. Cross-linking site in fibrinogen for α2-plasmin inhibitor. J Biol Chem 1986; 261: 15591-5.
    PubMedGoogle Scholar
  • 16 Sakata Y, Aoki N. Significance of cross-linking of α2-plasmin inhibitor to fibrin in inhibition of fibrinolysis and in hemostasis. J Clin Invest 1982; 69: 536-42.
    CrossrefPubMedGoogle Scholar
  • 17 Jansen JW, Haverkate F, Koopman J, Nieuwenhuis HK, Kluft C, Boschman TA. Influence of factor XIIIa activity on human whole blood clot lysis in vitro. Thromb Haemost 1987; 57: 171-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Reed 3rd GL, Matsueda GR, Haber E. Inhibition of clot-bound α2-antiplasmin enhances in vivo thrombolysis. Circulation 1990; 82: 164-8.
    CrossrefPubMedGoogle Scholar
  • 19 Aoki N, Sakata Y, Matsuda M, Tateno K. Fibrinolytic states in a patient with congenital deficiency of α2-plasmin inhibitor. Blood 1980; 55: 483-8.
    PubMedGoogle Scholar
  • 20 Aoki N. Genetic abnormalities of the fibrinolytic system. Semin Thromb Hemostas 1984; 10: 42-50.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Saito H. α2-plasmin inhibitor and its deficiency states. J Lab Clin Med 1988; 112: 671-8.
    PubMedGoogle Scholar
  • 22 Favier R, Aoki N, de Moerloose P. Congenital α2-plasmin inhibitor deficiencies: a review. Br J Haematol 2001; 114: 4-10.
    CrossrefPubMedGoogle Scholar
  • 23 Miura O, Hirosawa S, Kato A, Aoki N. Molecular basis for congenital deficiency of α2-plasmin inhibitor. A frameshift mutation leading to elongation of the deduced amino acid sequence. J Clin Invest 1989; 83: 1598-604.
    CrossrefPubMedGoogle Scholar
  • 24 Miura O, Sugahara Y, Aoki N. Hereditary α2-plasmin inhibitor deficiency caused by a transport-deficient mutation (α2-PI-Okinawa). Deletion of Glu137 by a trinucleotide deletion blocks intracellular transport. J Biol Chem 1989; 264: 18213-9.
    PubMedGoogle Scholar
  • 25 Miura O, Aoki N. Impaired secretion of mutant α2-plasmin inhibitor (α2PINara) from COS-7 and HepG2 cells: molecular and cellular basis for hereditary deficiency of α2-plasmin inhibitor. Blood 1990; 75: 1092-6.
    PubMedGoogle Scholar
  • 26 Lind B, Thorsen S. A novel missense mutation in the human plasmin inhibitor (α2-antiplasmin) gene associated with a bleeding tendency. Br J Haematol 1999; 107: 317-22.
    CrossrefPubMedGoogle Scholar
  • 27 Yoshinaga H, Hirosawa S, Chung DH, Miyasaka N, Aoki N, Favier R. A novel point mutation of the splicing donor site in the intron 2 of the plasmin inhibitor gene. Thromb Haemost 2000; 84: 307-11.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Holmes WE, Lijnen HR, Nelles L, Kluft C, Nieuwenhuis HK, Rijken DC, Collen D. α2-antiplasmin Enschede: alanine insertion and abolition of plasmin inhibitory activity. Science 1987; 238: 209-11.
    CrossrefPubMedGoogle Scholar
  • 29 Miles LA, Plow EF, Donnelly KJ, Hougie C, Griffin JH. A bleeding disorder due to deficiency of α2-antiplasmin. Blood 1982; 59: 1246-51.
    PubMedGoogle Scholar
  • 30 Mimuro J, Koike Y, Sumi Y, Aoki N. Monoclonal antibodies to discrete regions in α2-plasmin inhibitor. Blood 1987; 69: 446-53.
    PubMedGoogle Scholar
  • 31 Kimura S, Tamaki T, Aoki N. Acceleration of fibrinolysis by the N-terminal peptide of α2-plasmin inhibitor. Blood 1985; 66: 157-60.
    PubMedGoogle Scholar
  • 32 Lee KN, Tae WC, Jackson KW, Kwon SH, McKee PA. Characterization of wild-type and mutant α2-antiplasmins: fibrinolysis enhancement by reactive site mutant. Blood 1999; 94: 164-71.
    PubMedGoogle Scholar
  • 33 Sumi Y, Ichikawa Y, Nakamura Y, Miura O, Aoki N. Expression and characterization of pro α2-plasmin inhibitor. J Biochem (Tokyo) 1989; 106: 703-7.
    CrossrefPubMedGoogle Scholar
  • 34 Chung DH, Ohashi K, Watanabe M, Miyasaka N, Hirosawa S. Mannose trimming targets mutant α2-plasmin inhibitor for degradation by the proteasome. J Biol Chem 2000; 275: 4981-7.
    CrossrefPubMedGoogle Scholar