Hereditary Dysfunctional Protein C Molecules (Type II): Assay Characterization and Proposed Classification

Richard A Marlar; Dorothy M Adcock; Renée M Madden

doi:10.1055/s-0038-1645050

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1990; 63(03): 375-379
DOI: 10.1055/s-0038-1645050

Original Article

Schattauer GmbH Stuttgart

Hereditary Dysfunctional Protein C Molecules (Type II): Assay Characterization and Proposed Classification

Richard A Marlar

The Laboratory Service, Denver Veteran’s Administration Medical Center, Denver, Colorado, USA and the Department of Pathology, Department of Biochemistry, Biophysics and Genetics, Department of Pediatrics, University of Colorado Health Sciences Center, Denver, Colorado, USA

,

Dorothy M Adcock

The Laboratory Service, Denver Veteran’s Administration Medical Center, Denver, Colorado, USA and the Department of Pathology, Department of Biochemistry, Biophysics and Genetics, Department of Pediatrics, University of Colorado Health Sciences Center, Denver, Colorado, USA

,

Renée M Madden

The Laboratory Service, Denver Veteran’s Administration Medical Center, Denver, Colorado, USA and the Department of Pathology, Department of Biochemistry, Biophysics and Genetics, Department of Pediatrics, University of Colorado Health Sciences Center, Denver, Colorado, USA

› Author Affiliations

Abstract

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Summary

Protein C (PC) deficiency is among the increasing number of recognized causes of hereditary thrombotic disease. Two types of PC deficiency have been described: 1) Type I, which is characterized by a concomitant decrease in PC activity and antigen, and 2) Type II. characterized by disproportionately low activity compared to antigen (i.e. a dysfunctional molecule). To date, only a small number of Type II patients have been described. This study was undertaken to evaluate a number of dysfunctional PC molecules by comparing PC clotting and amidolytic activities with antigen levels. For these studies, an automated PTT-based clotting PC assay was developed. This assay was sensitive to 1% of a normal plasma pool, specific, accurate, and reproducible (± 12%). A good correlation (r = 0.918) of the clotting activity to antigen was found in normal individuals and Type I heterozygous and homozygous patients. To classify Type II PC deficient patients, the antigen, amidolytic and clotting PC levels were compared in ten affected families. The clotting activities were decreased in all affected members, whereas the antigen levels were within the normal limits. In four of the 10 families, the amidolytic activity was normal and similar to the antigen levels. This suggests that in certain families, defects in the PC molecule occur in regions not associated with amidolytic functions. From these studies, the molecular basis of Type II PC deficiency is varied and complex, involving different functional domains of the PC molecule. Therefore, we have suggested a nomenclature algorithm for Type II PC deficiency based on the location of the defect within the specific domains of the PC molecule.

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References

References
1 Clouse LH, Comp PC. The regulation of hemostasis: the protein C system. N Eng J Med 1986; 314: 1298-304
2 Esmon CT. The roles of protein C and thrombomodulin in the regulation of blood coagulation. J Biol Chem 1989; 264: 4743-6
3 Stenflo J. Structure and function of protein C. Sem Thromb Hemostas 1984; 10: 109-21
4 Esmon NL. Thrombomodulin. Sem Thromb Hemostas 1987; 13: 454-71
5 Marlar RA, Kleiss AJ, Griffin JH. Mechanism of action of human activated protein C, a thrombin-dependent anticoagulant enzyme. Blood 1982; 59: 1067-72
6 Walker FJ. Regulation of activated protein C by protein S. J Biol Chem 1981; 256: 11128-31
7 Brockmans AW, Conard J. Hereditary protein C deficiency. In: Protein C and Related Proteins Bertina R. (ed) Churchill Livingstone Publishers; New York: 1988: 160-81
8 Mannucci PM, Vigano S. Deficiencies of protein C. an inhibitor of blood coagulation. Lancet 1982; 2: 463-7
9 Marlar RA, Endres-Brooks J, Miller C. Serial studies of protein C and its inhibitor in patients with disseminated intravascular coagulation. Blood 1985; 66: 59-63
10 Marlar RA. Protein C in thromboembolic disease. Sem Thromb Hemostas 1985 11: 389-95
11 Marlar RA, Montgomery R, Brockmans A. Diagnosis and treatment of homozygous protein C-deficient children. J Ped 1989; 114: 528-35
12 Griffin JH, Evatt B, Zimmerman TS, Kleiss AJ, Wideman C. Deficiency of protein C in congenital thrombotic disease. J Clin Invest 1981; 68: 1370-3
13 Conlan MG, Bridges A, Williams E, Marlar RA. Familial type II protein C deficiency with Warfarin-induced skin necrosis and bilateral adrenal hemorrhage. Am J Hematol 1988; 29: 226-9
14 Gruppo RA, Francis RB, Marlar RA, Silberstein EB. Protein C deficiency resulting from possible double heterozygosity and its response to danazol. Blood 1988; 71: 370-4
15 Matsuda M, Sugo T, Sakata Y, Murayama H, Mimuro J, Tanabe S, Yoshitake S. A thrombotic state due to an abnormal protein C. N Eng J Med 1988; 319: 1265-8
16 Faioni EM, Esmon CT, Esmon NL, Mannucci PM. Isolation of an abnormal protein C molecule from the plasma of a patient with thrombotic diathesis. Blood 1988; 71: 940-6
17 Barbui T, Finazzi G, Mussoni L, Riganti M, Donati MB, Colucci M, Collen D. Hereditary dysfunctional protein C (protein C-Bcrgamo) and thrombosis. Lancet 1984; 2: 819
18 Sala N, Borrell M, Bauer KA, Vigano-D’Angelo S, Fontcuberta J, Felez J, Rutllant ML. Dysfunctional activated protein C (PC Cadiz) in a patient with thrombotic disease. Thromb Haemostas 1987; 57: 183-6
19 Comp PC, Nixon RR, Esmon CT. Determination of functional levels of protein C, an antithrombotic protein, using thrombin-throm-bomodulin complex. Blood 1984; 63: 15-21
20 Manco-Johnson MJ, Marlar RA, Jacobson LJ, Hays T, Warady BA. Severe protein C deficiency in newborn infants. J Ped 1988; 113: 359-63
21 Sala N, Owen WG, Collen D. A functional assay of protein C in human plasma. Blood 1984; 63: 671-5
22 Bertina RM, Brockmans AW, Krommenhock-van EC, van Wijngaarden A. The use of a functional and immunological assay for plasma protein C in the study of the heterogeneity of congenital protein C deficiency. Thromb Haemostas 1984; 51: 1-5
23 Francis RB. A simplified PTT-based protein C activity assay using thrombin-thrombomodulin complex. Thromb Res 1986; 37: 337-44
24 Martinoli JL, Stocker K. Fast functional protein C assay using Protac, a novel protein C activator. Thromb Res 1986; 43: 253-64
25 Mannucci PM, Boyer C, Tripodi A, Vigano-D’Angelo S, Wolf M, Valsecchi C, D’Angelo A, Meyer D, Larricu MJ. Multicenter comparison of five functional and two immunological assays for protein C. Thromb Haemostas 1987; 57: 44-8
26 Fair DS, Marlar RA. Biosynthesis and secretion of factor VII. protein C, protein S and the protein C inhibitor from a human hepatoma cell line. Blood 1986 67. 64-70
27 Esmon NL, Owen WG, Esmon CT. Isolation of a membrane-bound cofactor for thrombin-catalyzed activation of protein C. J Biol Chem 1982; 257: 859-64
28 Laurell CB. Electroimmunoassay. Scand J Clin Lab Invest 1977; 29: 21-32
29 Klein JD, Walker FJ. Purification of a protein C activator from the venom of the Southern Copperhead Snake (Agkistrodon contorthx). Biochemistry 1986; 25: 4175-9
30 Madden RM, Gill JC, Marlar RA. Protein C and protein S levels in acquired purpura fulminans. Br J Haematol, in press
31 Comp P. Hereditary disorders predisposing to thrombosis. In: Progress in Hemostasis and Thrombosis Coller BS. (ed). Grune and Stratton Inc; New York: 1986: 71-102
32 Mannucci PM, Tripodi A. Laboratory screening of inherited thrombotic syndromes. Thromb Haemostas 1987; 57: 247-51
33 Fair DS, Revak DJ, Hubbard JG, Girolami A. Isolation and characterization of the factor X-Friuli variant. Blood 1989; 73e: 2108-16
34 Ware J, Davis L, Frazier D, Bajaj SP, Stafford DW. Genetic defect responsible for the dysfunctional protein: Factor IX-Long Beach. Blood 1988; 72: 820-2
35 Sugo T, Persson U, Stenflo J. Protein C in bovine plasma after warfarin treatment. J Biol Chem 1985; 260: 10453-7
36 Krishnaswamy S, Williams EB, Mann KG. The binding of activated protein C to factors V and Va. J Biol Chem 1986; 261: 9684-93
37 Kidd VJ, Woo SL C. Molecular diagnosis of human genetic disease. Mod Cell Biol 1984; 3: 113-29
38 White GC, Shoemaker CB. Factor VIII gene and hemophilia A. Blood 1989; 73: 1-12
39 Geddes VA, Le Bonnicc BF, Louie GV, Brayer GD, Thompson AR, MacGillivary R TA. A moderate form of hemophilia B is caused by a novel mutation in the protease domain of factor IX-Vancouver. J Biol Chem 1989; 264: 4689-97
40 Sambrano JE, Jacobsonz LJ, Reeve EB, Manco-Johnson MJ, Hathaway WE. Abnormal antithrombin III with defective serine protease binding (antithrombin III “Denver”). J Clin Invest 1986; 77: 887-93
41 Holmberg L, Nilsson IM. Von Willebrand disease. Clin Haematol 1985; 14 (02) 461-88
42 Sas G. Classification of antithrombin III deficiencies – Has a new Tower of Babel been built. Thromb Haemostas 1988; 60: 530-1
43 Beckmann RJ, Schmidt RR, Santerrc FF, Plutzky J, Crabtree GR, Long GL. The stucture and evolution of a 461 amino acid human protein C precursor and its messenger RNA, based upon the DNA sequence of cloned human liver cDNAs. Nucleic Acid Res 1985; 13: 5233-47