Molecular Mechanism of Type I Congenital Heparin Cofactor (HC) II Deficiency Caused by a Missense Mutation at Reactive P2 Site: HC II Tokushima

Yasuhiko Kanagawa; Toshio Shigekiyo; Ken-ichi Aihara; Masashi Akaike; Hiroyuki Azuma; Toshio Matsumoto

doi:10.1055/s-0037-1612911

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2001; 85(01): 101-107
DOI: 10.1055/s-0037-1612911

Review Article

Schattauer GmbH

Molecular Mechanism of Type I Congenital Heparin Cofactor (HC) II Deficiency Caused by a Missense Mutation at Reactive P2 Site: HC II Tokushima

Autoren

Yasuhiko Kanagawa

¹First Department of Internal Medicine, University of Tokushima School of Medicine, Tokushima, Japan
Toshio Shigekiyo

²Tokushima Prefectural Central Hospital, Tokushima, Japan
Ken-ichi Aihara

¹First Department of Internal Medicine, University of Tokushima School of Medicine, Tokushima, Japan
Masashi Akaike

¹First Department of Internal Medicine, University of Tokushima School of Medicine, Tokushima, Japan
Hiroyuki Azuma

¹First Department of Internal Medicine, University of Tokushima School of Medicine, Tokushima, Japan
Toshio Matsumoto

¹First Department of Internal Medicine, University of Tokushima School of Medicine, Tokushima, Japan

Abstract

Summary

We found a 66-year-old Japanese patient with type I congenital heparin cofactor (HC) II deficiency manifesting multiple atherosclerotic lesions. To investigate the molecular pathogenesis of our patient, we performed sequencing analysis and expressed recombinant human wild-type and mutant HC II molecules in COS-1 and CHO-K1 cells. Sequencing analysis following amplification of each of all 5 exons and its flanking region showed a single C to T transition at nucleotide position 12,854 in exon 5, which changed a Pro⁴⁴³ codon (CCG) to Leu codon (CTG). Because this mutation generates a new Bbv I site, the Bbv I digestion pattern of the PCR-amplified exon 5 fragments from each family member was analyzed. In all cases, the patterns were consistent with the activities and antigen levels of plasma HC II in those members. Transient transfection, metabolic labeling and pulse-chase experiments followed by immunoprecipitation analysis showed that the recombinant mutant HC II molecules were secreted from COS-1 cells in reduced amounts compared with the wild-type, and that an enhanced intracellular association of the mutant molecules with a chaperone, GRP78/BiP, was observed in CHO-K1 cells. Northern blot analysis indicated that the mutant HC II mRNA was transcribed at a similar level as that of wild-type.

Immunohistochemical staining of the transfected cells revealed that COS-1 cells expressing the mutant HC II molecules were stained mainly in the perinuclear area. We conclude that the impaired secretion of the mutant HC II molecules, due to intracellular degradation, is the molecular pathogenesis of type I congenital HC II deficiency caused by a Pro⁴⁴³ to Leu mutation at reactive P2 site.

Keywords

Heparin cofactor II - thrombosis - thrombin - transfection - chaperone

Volltext

Referenzen

References
1 Tollefsen DM, Majerus PW, Blank MK. Heparin cofactor II. Purification and properties of a heparin-dependent inhibitor of thrombin in human plasma. J Biol Chem 1982; 257: 2162-9.
2 Ragg H. A new member of the plasma protease inhibitor gene family. Nucleic Acids Res 1986; 14: 1073-88.
3 Blinder MA, Marasa JC, Reynolds CH, Deaven LL, Tollefsen DM. Heparin cofactor II: cDNA sequence, chromosome localization, restriction length polymorphism, and expression in Escherichia coli. Biochemistry. 1988; 27: 752-9.
4 Tollefsen DM. Insight into the mechanism of action of heparin cofactor II. Thromb Haemost 1995; 74: 1209-14.
5 Parker KA, Tollefsen DM. The protease specificity of heparin cofactor II: Inhibition of thrombin generated during coagulation. J Biol Chem 1985; 260: 3501-5.
6 Church FC, Noyes CM, Griffith MJ. Inhibition of chymotrypsin by heparin cofactor II. Proc Natl Acad Sci USA 1985; 82: 6431-4.
7 Tollefsen DM, Pestka CA, Monafo WJ. Activation of heparin cofactor II by dermatan sulfate. J Biol Chem 1983; 258: 6713-6.
8 McGuire EA, Tollefsen DM. Activation of heparin cofactor II by fibroblasts and vascular smooth muscle cells. J Biol Chem 1987; 262: 169-75.
9 Sié P, Dupoy D, Pichon J, Boneu B. Constitutional heparin cofactor II deficiency associated with recurrent thrombosis. Lancet 1985; ii: 414-6.
10 Tran TH, Marbet GA, Duckert F. Association of hereditary heparin cofactor II deficiency with thrombosis. Lancet 1985; ii: 413-4.
11 Simioni P, Lazzaro AR, Coser E, Salmistraro G, Girolami A. Hereditary heparin cofactor II deficiency and thrombosis: report of six patients belonging to two separate kindreds. Blood Coag Fibrinol 1990; 1: 351-6.
12 Weisdorf DJ, Edson JR. Recurrent venous thrombosis associated with inherited deficiency of heparin cofactor II. Br J Haematol 1990; 77: 125-6.
13 Blinder MA, Andersson TR, Ablidgaard U, Tollefsen DM. Heparin cofactor II Oslo: Mutation of Arg-189 to His decreases the affinity for dermatan sulfate. J Biol Chem 1989; 264: 5128-33.
14 Matsuo T, Kario K, Sakamoto S, Yamada T, Miki T, Hirase T, Kobayashi H. Hereditary heparin cofactor II deficiency and coronary artery disease. Thromb Res 1992; 65: 495-505.
15 Awidi AS, Abu KM, Herzallah U, Abu RA, Shannak MM, Abu OT, Al TI, Anshasi B. Hereditary thrombophilia among 217 consecutive patients with thromboembolic disease in Jordan. Am J Hematol 1993; 44: 95-100.
16 Bernardi F, Legnani C, Micheletti F, Lunghi B, Ferraresi P, Palareti G, Biagi R, Marchetti G. A heparin cofactor II mutation (HC II Rimini) combined with factor V Leiden or type I Protein C deficiency in two unrelated thrombophilic subjects. Thromb Haemost 1996; 76: 505-9.
17 Villa P, Aznar J, Vaya A, Espana F, Ferrando F, Mira Y, Estelles A. Hereditary homozygous heparin cofactor II deficiency and the risk of developing venous thrombosis. Thromb Haemost 1999; 82: 1011-4.
18 Kondo S, Tokunaga F, Kario K, Matsuo T, Koide T. Molecular and cellular basis for type I heparin cofactor II deficiency (Heparin cofactor II Awaji). Blood 1996; 87: 1006-12.
19 Bertina RM, van der Linden IK, Engesser L, Muller HP, Brommer JP. Hereditary heparin cofactor II deficiency and the risk of development of thrombosis. Thromb Haemost 1987; 57: 196-200.
20 Azuma H, Uno Y, Shigekiyo T, Saito S. Congenital plasminogen deficiency caused by a Ser 572 to Pro mutation. Blood 1993; 82: 475-80.
21 Herzog R, Lutz S, Blin N, Marasa JC, Blinder MA, Tollefsen DM. Complete nucleotide sequence of the gene for human heparin cofactor II and mapping to chromosomal band 22q11. Biochemistry 1991; 30: 1350-7.
22 Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 1979; 77: 1513-23.
23 Ragg H, Preibisch G. Structure and expression of the gene coding for the human serpin hLS2. J Biol Chem 1988; 263: 12129-34.
24 Lane DA, Bayston T, Olds RJ, Fitches AC, Cooper DN, Millar DS, Jochmans K, Perry DJ, Okajima K, Thein SL, Emmerich J. Antithrombin Mutation Database: 2nd (1997) Update. Thromb Haemost 1997; 77: 197-211.
25 Kuznetsov G, Nigam SK. Folding of secretory and membrane protein. N Eng J Med 1998; 339: 1688-95.
26 Zhang GS, Mehringer JH, Van Deerlin VM, Kozak CA, Tollefsen DM. Murine heparin cofactor II: purification, cDNA sequence, expression, and gene structure. Biochemistry 1994; 33: 3632-42.
27 Westrup D, Ragg H. Secondary thrombin-binding site, glycosaminoglycan binding domain and reactive center region of leuserpin-2 are strongly conserved in mammalian species. Biochim Biophys Acta 1994; 1217: 93-6.
28 Sheffield WP, Schuyler PD, Blajchman MA. Molecular cloning and expression of rabbit heparin cofactor II: A plasma thrombin inhibitor highly conserved between species. Thromb Haemost 1994; 71: 778-82.
29 Colwel NS, Tollefsen DM. Isolation of frog and chicken cDNA encoding heparin cofactor II. Thromb Haemost 1998; 80: 784-90.
30 Huber R, Carrell RW. Implications of the three-dimensional structure of α₁-antitrypsin for structure and function of serpins. Biochemistry 1989; 28: 8951-66.
31 Blajchman MA, Fernandez-Rachubinsky F, Sheffield WP, Austin RC, Schulman S. Antithrombin III Stockholm: A codon 392 (Gly to Asp) mutation with normal heparin binding and impaired serine protease reactivity. Blood 1992; 79: 1428-34.
32 Coughlin SR, Vu T-KH, Hung DT, Wheaton VI. Characterization of a functional thrombin receptor. Issues and opportunities. J Clin Invest 1992; 89: 351-5.
33 Ross R. The pathogenesis of atherosclerosis – an update. N Eng J Med 1986; 314: 488-500.
34 Fingerle J, Johnson R, Clows AW, Majesky MW, Reidy MA. Role of platelets in smooth muscle cell proliferation and migration after vascular injury in rat carotid artery. Proc Natl Acad Sci 1989; 86: 8412-6.
35 Nelken NA, Soifer SJ, O’Keefe J, Vu TK, Charo IF, Coughlin SR. Thrombin receptor expression in normal and atherosclerotic human arteries. J Clin Invest 1992; 90: 1614-21.
36 Hiramoto SA, Cunningham DD. Effects of fibroblasts and endothelial cells on inactivation of target proteases by protease nexin-1, heparin cofactor II and C1-inhibitor. J Cell Biochem 1988; 36: 199-207.
37 Riessen R, Isner JM, Blessing E, Loushin C, Nikol S, Wight TN. Regional differences in the distribution of the proteoglycans biglycan and decorin in the extracellular matrix of atherosclerotic and restenotic human coronary arteries. Am J Pthol 1994; 144: 962-74.