Prothrombin Scranton: Substitution of an Amino Acid Residue Involved in the Binding of Na⁺ (LYS-556 to THR) Leads to Dysprothrombinemia

Summary

Several members of a family from Scranton, Pennsylvania were identified to have normal levels of prothrombin antigen while their prothrombin clotting activity was approximately 50% of normal. There has been no previous history of bleeding or other clinical manifestations in this family. The genomic DNA from the proband was amplified for all exons in the prothrombin gene and analyzed by single strand conformation polymorphism (SSCP)/heteroduplex analysis followed by DNA sequence analysis and restriction enzyme digestion. A mutation at nucleotide 20040 in exon 14 was identified and confirmed by restriction enzyme digestion. This mutation results in the substitution of Thr for Lys at amino acid 556. Amino acid 556 has been reported as one of the key residues for the binding of Na⁺ in the thrombin portion of the protein.

Keywords

Prothrombin - dysprothrombinemia - abnormal prothrombin - Na⁺ binding - SSCP/heteroduplex analysis

• References

• 1 Davie EW, Fujikawa K, Kisiel W. The coagulation cascade: initiation, maintenance, and regulation. Biochemistry 1991; 30: 10363-70.
• 2 Di Cera E, Guinto ER, Vindigni A, Dang QD, Ayala YM, Wuyi M, Tulinsky A. The Na⁺ binding site of thrombin. J Biol Chem 1995; 270: 22089-92.
• 3 Dang QD, Sabetta M, Di Cera E. Selective loss of fibrinogen clotting in a loop-less thrombin. J Biol Chem 1997; 272: 19649-51.
• 4 Dang QD, Enriqueta RG, Di Cera E. Rational engineering of activity and specificity in a serine protease. Nature Biotech 1997; 15: 146-9.
• 5 Sun WY, Witte D, Degen JL, Colbert M, Burkart M, Holmback K, Xiao Q, Bugge T, Degen SJF. Prothrombin deficiency results in embryonic and neonatal lethality in mice. Proc Natl Acad Sci USA 1998; 95: 7597-602.
• 6 Xue J, Wu Q, Westfield LA, Tuley EA, Lu D, Zhang Q, Shim K, Zheng X, Sadler JE. Incomplete embryonic lethality and fatal neonatal hemorrhage caused by prothrombin deficiency in mice. Proc Natl Acad Sci USA 1998; 95: 7603-7.
• 7 Degen SJF, McDowell SA, Sparks LM, Scharrer I. Prothrombin Frankfurt: a dysfunctional prothrombin characterized by substitution of Glu-466 by Ala. Thromb Haemost 1995; 73: 203-9.
• 8 O’Marcaigh AS, Nichols WL, Hassinger NL, Mullins JD, Mallouh AA, Gilchrist GS, Owen WG. Genetic analysis and functional characterization of prothrombin Corpus Christi (Arg382-Cys), Dhahran (Arg271-His) and hypoprothrombinemia. Blood 1996; 88: 2611-8.
• 9 Sun WM, Burkart MC, Holahan JR, Degen SJF. Prothrombin San Antonio: a single amino acid substitution at a factor Xa activation site (Arg 320 to His) results in dysprothrombinemia. Blood 2000; 95 (Suppl. 02) 711-3.
• 10 Sun WY, Ruiz-Saez A, Burkart MC, Bosch N, Degen SJF. Prothrombin Carora: hypoprothrombinemia caused by substitution of Tyr-44 by Cys. British J. Haem 1999; 105: 670-2.
• 11 Degen SJF. Prothrombin. In: Mol. Biol. of Thrombosis and Hemostasis. Roberts H, High K. eds. New York: Marcel Dekker, Inc; 1995: 75-99.
• 12 Degen SJF, Davie EW. Nucleotide sequence of the gene for human prothrombin. Biochemistry 1987; 26: 6165-77.
• 13 Morishita E, Saito M, Kumabashiri I, Asakura H, Matsuda T, Yamaguchi K. Prothrombin Himi: A compound heterozygote for two dysfunctional prothrombin molecules (Met-337-Thr and Arg-388-His). Blood 1992; 80: 2275-80.
• 14 James HL, Kim DJ, Zheng DQ, Girolami A. Prothrombin Padua I: Incomplete activation due to an amino acid substitution at a factor Xa cleavage site. Blood Coagulation and Fibrinolysis 1994; 5: 841-4.
• 15 Banfield DK, MacGillivray RTA. Partial characterization of vertebrate prothrombin cDNAs: amplification and sequence analysis of the B chain of thrombin from nine different species. Proc Natl Acad Sci 1992; 89: 2779-83.