Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Aug;83(16):5803–5807. doi: 10.1073/pnas.83.16.5803

Molecular basis of hemophilia B: a defective enzyme due to an unprocessed propeptide is caused by a point mutation in the factor IX precursor.

D L Diuguid, M J Rabiet, B C Furie, H A Liebman, B Furie
PMCID: PMC386383  PMID: 3461460

Abstract

A mutant factor IX, designated factor IXCambridge, was isolated from a patient with hemophilia B. This protein includes an 18-residue propeptide attached to the NH2 terminus of factor IX. A point mutation at residue -1, from an arginine to a serine, precludes cleavage of the propeptide by a processing protease and interferes with gamma-carboxylation of the factor IX, indicating the importance of the leader sequence in substrate recognition by the vitamin K-dependent carboxylase. This represents an example of an enzyme defect due to the presence of a point mutation in a precursor protein (preproenzyme) that is the cause of a human hereditary disease. This defect will serve as a prototype for understanding the molecular basis of some forms of hemophilia and other hereditary enzyme deficiencies.

Full text

PDF
5806

Images in this article

5804Image
on p.5804

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abdo Y., Rousseaux J., Dautrevaux M. Proalbumin Lille, a new variant of human serum albumin. FEBS Lett. 1981 Aug 31;131(2):286–288. doi: 10.1016/0014-5793(81)80386-9. [DOI] [PubMed] [Google Scholar]
  2. Bajaj S. P. Cooperative Ca2+ binding to human factor IX. Effects of Ca2+ on the kinetic parameters of the activation of factor IX by factor XIa. J Biol Chem. 1982 Apr 25;257(8):4127–4132. [PubMed] [Google Scholar]
  3. Barenholz Y., Gibbes D., Litman B. J., Goll J., Thompson T. E., Carlson R. D. A simple method for the preparation of homogeneous phospholipid vesicles. Biochemistry. 1977 Jun 14;16(12):2806–2810. doi: 10.1021/bi00631a035. [DOI] [PubMed] [Google Scholar]
  4. Barwick R. C., Jones R. T., Head C. G., Shih M. F., Prchal J. T., Shih D. T. Hb Long Island: a hemoglobin variant with a methionyl extension at the NH2 terminus and a prolyl substitution for the normal histidyl residue 2 of the beta chain. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4602–4605. doi: 10.1073/pnas.82.14.4602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bentley A. K., Rees D. J., Rizza C., Brownlee G. G. Defective propeptide processing of blood clotting factor IX caused by mutation of arginine to glutamine at position -4. Cell. 1986 May 9;45(3):343–348. doi: 10.1016/0092-8674(86)90319-3. [DOI] [PubMed] [Google Scholar]
  6. Blanchard R. A., Furie B. C., Jorgensen M., Kruger S. F., Furie B. Acquired vitamin K-dependent carboxylation deficiency in liver disease. N Engl J Med. 1981 Jul 30;305(5):242–248. doi: 10.1056/NEJM198107303050502. [DOI] [PubMed] [Google Scholar]
  7. Blanchard R. A., Furie B. C., Kruger S. F., Waneck G., Jorgensen M. J., Furie B. Immunoassays of human prothrombin species which correlate with functional coagulant activities. J Lab Clin Med. 1983 Feb;101(2):242–255. [PubMed] [Google Scholar]
  8. Bloom J. W., Nesheim M. E., Mann K. G. Phospholipid-binding properties of bovine factor V and factor Va. Biochemistry. 1979 Oct 2;18(20):4419–4425. doi: 10.1021/bi00587a023. [DOI] [PubMed] [Google Scholar]
  9. Blouquit Y., Arous N., Lena D., Delanoe-Garin J., Lacombe C., Bardakdjian J., Vovan L., Orsini A., Rosa J., Galacteros F. Hb Marseille [alpha 2 beta 2 N methionyl-2 (NA2) His----Pro]: a new beta chain variant having an extended N-terminus. FEBS Lett. 1984 Dec 10;178(2):315–318. doi: 10.1016/0014-5793(84)80624-9. [DOI] [PubMed] [Google Scholar]
  10. Boissel J. P., Kasper T. J., Shah S. C., Malone J. I., Bunn H. F. Amino-terminal processing of proteins: hemoglobin South Florida, a variant with retention of initiator methionine and N alpha-acetylation. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8448–8452. doi: 10.1073/pnas.82.24.8448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Borowski M., Furie B. C., Furie B. Distribution of gamma-carboxyglutamic acid residues in partially carboxylated human prothrombins. J Biol Chem. 1986 Feb 5;261(4):1624–1628. [PubMed] [Google Scholar]
  12. Borowski M., Furie B. C., Goldsmith G. H., Furie B. Metal and phospholipid binding properties of partially carboxylated human prothrombin variants. J Biol Chem. 1985 Aug 5;260(16):9258–9264. [PubMed] [Google Scholar]
  13. Brennan S. O., Carrell R. W. A circulating variant of human proalbumin. Nature. 1978 Aug 31;274(5674):908–909. doi: 10.1038/274908a0. [DOI] [PubMed] [Google Scholar]
  14. Choo K. H., Gould K. G., Rees D. J., Brownlee G. G. Molecular cloning of the gene for human anti-haemophilic factor IX. Nature. 1982 Sep 9;299(5879):178–180. doi: 10.1038/299178a0. [DOI] [PubMed] [Google Scholar]
  15. DiScipio R. G., Davie E. W. Characterization of protein S, a gamma-carboxyglutamic acid containing protein from bovine and human plasma. Biochemistry. 1979 Mar 6;18(5):899–904. doi: 10.1021/bi00572a026. [DOI] [PubMed] [Google Scholar]
  16. Giannelli F., Choo K. H., Rees D. J., Boyd Y., Rizza C. R., Brownlee G. G. Gene deletions in patients with haemophilia B and anti-factor IX antibodies. Nature. 1983 May 12;303(5913):181–182. doi: 10.1038/303181a0. [DOI] [PubMed] [Google Scholar]
  17. Hauschka P. V. Quantitative determination of gamma-carboxyglutamic acid in proteins. Anal Biochem. 1977 May 15;80(1):212–223. doi: 10.1016/0003-2697(77)90640-6. [DOI] [PubMed] [Google Scholar]
  18. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [PubMed] [Google Scholar]
  19. Kurachi K., Davie E. W. Isolation and characterization of a cDNA coding for human factor IX. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6461–6464. doi: 10.1073/pnas.79.21.6461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Liebman H. A., Limentani S. A., Furie B. C., Furie B. Immunoaffinity purification of factor IX (Christmas factor) by using conformation-specific antibodies directed against the factor IX-metal complex. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3879–3883. doi: 10.1073/pnas.82.11.3879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Long G. L., Belagaje R. M., MacGillivray R. T. Cloning and sequencing of liver cDNA coding for bovine protein C. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5653–5656. doi: 10.1073/pnas.81.18.5653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McMullen B. A., Fujikawa K., Kisiel W. The occurrence of beta-hydroxyaspartic acid in the vitamin K-dependent blood coagulation zymogens. Biochem Biophys Res Commun. 1983 Aug 30;115(1):8–14. doi: 10.1016/0006-291x(83)90961-0. [DOI] [PubMed] [Google Scholar]
  24. Morita T., Isaacs B. S., Esmon C. T., Johnson A. E. Derivatives of blood coagulation factor IX contain a high affinity Ca2+-binding site that lacks gamma-carboxyglutamic acid. J Biol Chem. 1984 May 10;259(9):5698–5704. [PubMed] [Google Scholar]
  25. Nelsestuen G. L., Lim T. K. Equilibria involved in prothrombin- and blood-clotting factor X-membrane binding. Biochemistry. 1977 Sep 20;16(19):4164–4171. doi: 10.1021/bi00638a005. [DOI] [PubMed] [Google Scholar]
  26. Nelsestuen G. L., Resnick R. M., Wei G. J., Pletcher C. H., Bloomfield V. A. Metal ion interactions with bovine prothrombin and prothrombin fragment 1. Stoichiometry of binding, protein self-association, and conformational change induced by a variety of metal ions. Biochemistry. 1981 Jan 20;20(2):351–358. doi: 10.1021/bi00505a019. [DOI] [PubMed] [Google Scholar]
  27. Nelsestuen G. L., Zytkovicz T. H., Howard J. B. The mode of action of vitamin K. Identification of gamma-carboxyglutamic acid as a component of prothrombin. J Biol Chem. 1974 Oct 10;249(19):6347–6350. [PubMed] [Google Scholar]
  28. Nemerson Y., Furie B. Zymogens and cofactors of blood coagulation. CRC Crit Rev Biochem. 1980;9(1):45–85. doi: 10.3109/10409238009105472. [DOI] [PubMed] [Google Scholar]
  29. Noyes C. M., Griffith M. J., Roberts H. R., Lundblad R. L. Identification of the molecular defect in factor IX Chapel Hill: substitution of histidine for arginine at position 145. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4200–4202. doi: 10.1073/pnas.80.14.4200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pan L. C., Price P. A. The propeptide of rat bone gamma-carboxyglutamic acid protein shares homology with other vitamin K-dependent protein precursors. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6109–6113. doi: 10.1073/pnas.82.18.6109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prendergast F. G., Mann K. G. Differentiation of metal ion-induced transitions of prothrombin fragment 1. J Biol Chem. 1977 Feb 10;252(3):840–850. [PubMed] [Google Scholar]
  32. Rees D. J., Rizza C. R., Brownlee G. G. Haemophilia B caused by a point mutation in a donor splice junction of the human factor IX gene. Nature. 1985 Aug 15;316(6029):643–645. doi: 10.1038/316643a0. [DOI] [PubMed] [Google Scholar]
  33. Robbins D. C., Blix P. M., Rubenstein A. H., Kanazawa Y., Kosaka K., Tager H. S. A human proinsulin variant at arginine 65. Nature. 1981 Jun 25;291(5817):679–681. doi: 10.1038/291679a0. [DOI] [PubMed] [Google Scholar]
  34. Shibasaki Y., Kawakami T., Kanazawa Y., Akanuma Y., Takaku F. Posttranslational cleavage of proinsulin is blocked by a point mutation in familial hyperproinsulinemia. J Clin Invest. 1985 Jul;76(1):378–380. doi: 10.1172/JCI111973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stenflo J., Fernlund P., Egan W., Roepstorff P. Vitamin K dependent modifications of glutamic acid residues in prothrombin. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2730–2733. doi: 10.1073/pnas.71.7.2730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stenflo J., Suttie J. W. Vitamin K-dependent formation of gamma-carboxyglutamic acid. Annu Rev Biochem. 1977;46:157–172. doi: 10.1146/annurev.bi.46.070177.001105. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES