Hypofibrinogenaemia associated with a novel heterozygous γ289 Ala→Val substitution (fibrinogen Dorfen)

Amy Dear; Stephen O. Brennan; Carl-Erik Dempfle; Werner Kirschstein; Peter M. George

doi:10.1160/TH04-07-0409

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2004; 92(06): 1291-1295
DOI: 10.1160/TH04-07-0409

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Hypofibrinogenaemia associated with a novel heterozygous γ289 Ala→Val substitution (fibrinogen Dorfen)

Amy Dear

¹Molecular Pathology Laboratory, Christchurch School of Medicine and Health Sciences, University of Otago, Christchurch, New Zealand

,

Stephen O. Brennan

¹Molecular Pathology Laboratory, Christchurch School of Medicine and Health Sciences, University of Otago, Christchurch, New Zealand

,

Carl-Erik Dempfle

²First Department of Medicine, University Hospital of Mannheim, Mannheim, Germany

,

Werner Kirschstein

²First Department of Medicine, University Hospital of Mannheim, Mannheim, Germany

,

Peter M. George

¹Molecular Pathology Laboratory, Christchurch School of Medicine and Health Sciences, University of Otago, Christchurch, New Zealand

› Institutsangaben

Abstract

Summary

The molecular basis of hypofibrinogenaemia was investigated in a 34-year-old woman and her 10-year-old daughter. DNA sequencing revealed a single heterozygous GCC→GTC transition in exon 8 of the fibrinogen γ gene in both subjects, predicting a novel γ289 Ala→Val substitution. Examination of fibrinogen γ chains by electrospray ionization mass spectrometry failed to detect the variant chain in plasma fibrinogen. Further evidence for its non-expression came from tryptic peptide mapping. The mutation predicts a mass increase of 28 Da in peptide T32, but only the normal (M+2H) ion was detected at 1418 m/z in the proposita. Our finding that γ²⁸⁹ is an important determinant of plasma fibrinogen levels highlights the role of mutational analysis in defining structurally important regions of the fibrinogen molecule. This case suggests that the highly conserved Ala289 is important in maintaining structure of the “a” polymerization site via hydrogen bonding to Thr³⁷¹.

Keywords

Fibrinogen/fibrin - inherited coagulation disorders - gene mutations - protein structure/folding

Volltext

Referenzen

References
1 Henschen AH, McDonagh J. Fibrinogen, fibrin and factor XIII. In: Blood Coagulation. Vol 13. Zwaal RFA, Hemker HC. eds. Amsterdam: Elsevier Science Publishers BV; 1986: 171-241.
2 Doolittle RF. The molecular biology of fibrin. In: The Molecular Basis of Blood Diseases. Stamatoyannopoulos G, Nienhuis AW, Majerus PW, Varmus H. eds. Philadelphia: WB Saunders Company; 1994: 701-26.
3 Marchant RE, Barb MD, Shainoff R. et al. Three dimensional structure of human fibrinogen under aqueous conditions visualised by atomic force microscopy. Thromb Haemost 1997; 77: 1048-51.
4 Spraggon G, Everse SJ, Doolittle RF. Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin. Nature 1997; 389: 455-62.
5 Chen R, Doolittle RF. γ-γ cross-linking sites in human and bovine fibrinogen. Biochemistry 1971; 10: 4486-91.
6 Huang S, Mulvhill ER, Farrell DH. et al. Biosynthesis of human fibrinogen. Subunit interactions and potential intermediates in assembly. J Biol Chem 1993; 268: 8919-26.
7 Huang S, Cao Z, Chung DW. et al. The role of βγ and αγ complexes in the assembly of human fibrinogen. J Biol Chem 1996; 271: 27942-7.
8 Brennan SO, Hammonds B, George PM. Aberrant hepatic processing causes removal of activation peptide and primary polymerisation site from fibrinogen Canterbury (Aα20Val→ Asp). J Clin Invest 1995; 96: 2854-8.
9 Everse SJ, Spraggon G, Doolittle RF. A threedimensional consideration of variant human fibrinogens. Thromb Haemost 1998; 80: 1-9.
10 Cote HC, Lord ST, Pratt KP. γ-Chain dysfibrinogenemias: molecular structure-function relationships of naturally occurring mutations in the gamma chain of human fibrinogen. Blood 1998; 92: 2195-212.
11 Brennan SO, Fellowes AP, George PM. Molecular mechanisms of hypo-and afibrinogenemia. Ann NY Acad Sci 2001; 936: 91-100.
12 Maghzal GJ, Brennan SO, Homer VM. et al. The molecular mechanisms of congenital hypofibrinogenaemia. Cell Mol Life Sci 2004; 61: 1427-38.
13 Ciulla TA, Sklar RM, Hauser SL. A simple method for DNA purification from peripheral blood. Anal Biochem 1988; 174: 485-8.
14 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
15 Brennan SO. Electrospray ionisation analysis of human fibrinogen. Thromb Haemost 1997; 78: 1055-8.
16 Yee VC, Pratt KP, Cote HCF. et al. Crystal structure of a 30 kDa C-terminal fragment from the γ chain of human fibrinogen. Structure 1997; 05: 125-38. (supplementary material)
17 Brennan SO, Wyatt JM, Fellowes AP. et al. γ371 Thr→Ile substitution in the fibrinogen γD domain causes hypofibrinogenaemia. Biochim Biophys Acta 2001; 1550: 183-8.
18 Meyer M, Franke K, Richter W. et al. New molecular defects in the γ subdomain of fibrinogen D-domain in four cases of (hypo) dysfibrinogenemia: fibrinogen variants Hannover VI, Homburg VII, Stuttgart and Suhl. Thromb Haemost 2003; 89: 637-46.
19 Brennan SO, Wyatt JM, Medicina D. et al. Fibrinogen Brescia: Hepatic endoplasmic reticulum storage and hypofibrinogenemia because of a γ284 Gly→Arg mutation. Am J Pathol 2000; 157: 189-96.
20 Brennan SO, Maghzal G, Shneider BL. et al. Novel fibrinogen γ375 Arg→Trp mutation (fibrinogen Aguadilla) causes hepatic endoplasmic reticulum storage and hypofibrinogenemia. Hepatology 2002; 36: 652-8.