Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1992 May;89(5):1537–1543. doi: 10.1172/JCI115746

Met 358 to Arg mutation of alpha 1-antitrypsin associated with protein C deficiency in a patient with mild bleeding tendency.

D Vidaud 1, J Emmerich 1, M Alhenc-Gelas 1, J Yvart 1, J N Fiessinger 1, M Aiach 1
PMCID: PMC443026  PMID: 1569192

Abstract

The molecular defect responsible for a dramatic prolongation of all standard clotting tests discovered in a 15-yr-old boy has been identified. Initial investigations revealed the presence of an activated Factor X (Factor Xa) and thrombin inhibitor which copurified with alpha 1-antitrypsin (alpha 1-AT), thereby suggesting the occurrence of an alpha 1-AT variant similar to alpha 1-AT Pittsburgh. This was confirmed by dot-blot analysis and direct sequencing after amplification by the polymerase chain reaction. A G to T transition at nucleotide 10038 results in the substitution of Met to an Arg, converting alpha 1-AT into an Arg-Ser protease inhibitor (serpin) that inhibited thrombin and Factor Xa more effectively than antithrombin III. Surprisingly, there was no bleeding history in the proband. The common mutation Z, which may explain a reduced expression of the allele bearing the Arg 358 Met mutation, was not observed in the propositus' DNA. To exclude the presence of another mutation, the coding regions and intron/exon junctions were sequenced. No other mutation was found. Recently, the patient experienced his first hemorrhagic episode at the age of 17. The level of the abnormal inhibitor had increased twofold 2 mo before. The large decrease in protein C concentration may account for the mild bleeding tendency in this case, despite the presence of the alpha 1-AT Pittsburgh mutation. An abnormal protein C pattern was observed in patient's plasma, suggesting that the circulating deficiency might be due to a deleterious effect of the abnormal inhibitor on both intracellular processing and catabolism of protein C.

Full text

PDF
1537

Images in this article

1539Image
on p.1539
1540Image
on p.1540
1540Image
on p.1540
1541Image
on p.1541
1541Image
on p.1541
1541Image
on p.1541
1541Image
on p.1541
1541Image
on p.1541
1541Image
on p.1541
1541Image
on p.1541
1541Image
on p.1541

Selected References

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

  1. Barr P. J. Mammalian subtilisins: the long-sought dibasic processing endoproteases. Cell. 1991 Jul 12;66(1):1–3. doi: 10.1016/0092-8674(91)90129-m. [DOI] [PubMed] [Google Scholar]
  2. Bertina R. M., Broekmans A. W., Krommenhoek-van Es C., van Wijngaarden A. The use of a functional and immunologic assay for plasma protein C in the study of the heterogeneity of congenital protein C deficiency. Thromb Haemost. 1984 Feb 28;51(1):1–5. [PubMed] [Google Scholar]
  3. Brennan S. O., Owen M. C., Boswell D. R., Lewis J. H., Carrell R. W. Circulating proalbumin associated with a variant proteinase inhibitor. Biochim Biophys Acta. 1984 Nov 6;802(1):24–28. doi: 10.1016/0304-4165(84)90029-1. [DOI] [PubMed] [Google Scholar]
  4. Brennan S. O., Peach R. J. Calcium-dependent KEX2-like protease found in hepatic secretory vesicles converts proalbumin to albumin. FEBS Lett. 1988 Feb 29;229(1):167–170. doi: 10.1016/0014-5793(88)80819-6. [DOI] [PubMed] [Google Scholar]
  5. Carrell R. W., Aulak K. S., Owen M. C. The molecular pathology of the serpins. Mol Biol Med. 1989 Feb;6(1):35–42. [PubMed] [Google Scholar]
  6. Crystal R. G. Alpha 1-antitrypsin deficiency, emphysema, and liver disease. Genetic basis and strategies for therapy. J Clin Invest. 1990 May;85(5):1343–1352. doi: 10.1172/JCI114578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Esmon C. T., Owen W. G. Identification of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2249–2252. doi: 10.1073/pnas.78.4.2249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Esmon C. T. The roles of protein C and thrombomodulin in the regulation of blood coagulation. J Biol Chem. 1989 Mar 25;264(9):4743–4746. [PubMed] [Google Scholar]
  9. Esmon N. L., Owen W. G., Esmon C. T. Isolation of a membrane-bound cofactor for thrombin-catalyzed activation of protein C. J Biol Chem. 1982 Jan 25;257(2):859–864. [PubMed] [Google Scholar]
  10. Foster D. C., Holly R. D., Sprecher C. A., Walker K. M., Kumar A. A. Endoproteolytic processing of the human protein C precursor by the yeast Kex2 endopeptidase coexpressed in mammalian cells. Biochemistry. 1991 Jan 15;30(2):367–372. doi: 10.1021/bi00216a009. [DOI] [PubMed] [Google Scholar]
  11. Foster D. C., Sprecher C. A., Holly R. D., Gambee J. E., Walker K. M., Kumar A. A. Endoproteolytic processing of the dibasic cleavage site in the human protein C precursor in transfected mammalian cells: effects of sequence alterations on efficiency of cleavage. Biochemistry. 1990 Jan 16;29(2):347–354. doi: 10.1021/bi00454a007. [DOI] [PubMed] [Google Scholar]
  12. Greffe B. S., Manco-Johnson M. J., Marlar R. A. Molecular forms of human protein C: comparison and distribution in human adult plasma. Thromb Haemost. 1989 Nov 24;62(3):902–905. [PubMed] [Google Scholar]
  13. Griffin J. H., Evatt B., Zimmerman T. S., Kleiss A. J., Wideman C. Deficiency of protein C in congenital thrombotic disease. J Clin Invest. 1981 Nov;68(5):1370–1373. doi: 10.1172/JCI110385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heeb M. J., Bischoff R., Courtney M., Griffin J. H. Inhibition of activated protein C by recombinant alpha 1-antitrypsin variants with substitution of arginine or leucine for methionine358. J Biol Chem. 1990 Feb 5;265(4):2365–2369. [PubMed] [Google Scholar]
  15. Heeb M. J., Griffin J. H. Physiologic inhibition of human activated protein C by alpha 1-antitrypsin. J Biol Chem. 1988 Aug 25;263(24):11613–11616. [PubMed] [Google Scholar]
  16. Heeb M. J., Griffin J. H. Physiologic inhibition of human activated protein C by alpha 1-antitrypsin. J Biol Chem. 1988 Aug 25;263(24):11613–11616. [PubMed] [Google Scholar]
  17. Lewis J. H., Iammarino R. M., Spero J. A., Hasiba U. Antithrombin Pittsburgh: an alpha1-antitrypsin variant causing hemorrhagic disease. Blood. 1978 Jan;51(1):129–137. [PubMed] [Google Scholar]
  18. Long G. L., Chandra T., Woo S. L., Davie E. W., Kurachi K. Complete sequence of the cDNA for human alpha 1-antitrypsin and the gene for the S variant. Biochemistry. 1984 Oct 9;23(21):4828–4837. doi: 10.1021/bi00316a003. [DOI] [PubMed] [Google Scholar]
  19. Misumi Y., Oda K., Fujiwara T., Takami N., Tashiro K., Ikehara Y. Functional expression of furin demonstrating its intracellular localization and endoprotease activity for processing of proalbumin and complement pro-C3. J Biol Chem. 1991 Sep 5;266(25):16954–16959. [PubMed] [Google Scholar]
  20. Misumi Y., Ohkubo K., Sohda M., Takami N., Oda K., Ikehara Y. Intracellular processing of complement pro-C3 and proalbumin is inhibited by rat alpha 1-protease inhibitor variant (Met352----Arg) in transfected cells. Biochem Biophys Res Commun. 1990 Aug 31;171(1):236–242. doi: 10.1016/0006-291x(90)91382-3. [DOI] [PubMed] [Google Scholar]
  21. Owen M. C., Brennan S. O., Lewis J. H., Carrell R. W. Mutation of antitrypsin to antithrombin. alpha 1-antitrypsin Pittsburgh (358 Met leads to Arg), a fatal bleeding disorder. N Engl J Med. 1983 Sep 22;309(12):694–698. doi: 10.1056/NEJM198309223091203. [DOI] [PubMed] [Google Scholar]
  22. Petersen K. B., Kølvraa S., Bolund L., Petersen G. B., Koch J., Gregersen N. Detection of alpha 1-antitrypsin genotypes by analysis of amplified DNA sequences. Nucleic Acids Res. 1988 Jan 11;16(1):352–352. doi: 10.1093/nar/16.1.352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scott C. F., Carrell R. W., Glaser C. B., Kueppers F., Lewis J. H., Colman R. W. Alpha-1-antitrypsin-Pittsburgh. A potent inhibitor of human plasma factor XIa, kallikrein, and factor XIIf. J Clin Invest. 1986 Feb;77(2):631–634. doi: 10.1172/JCI112346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Suzuki K., Nishioka J., Hashimoto S. Protein C inhibitor. Purification from human plasma and characterization. J Biol Chem. 1983 Jan 10;258(1):163–168. [PubMed] [Google Scholar]
  27. Travis J., Matheson N. R., George P. M., Carrell R. W. Kinetic studies on the interaction of alpha 1-proteinase inhibitor (Pittsburgh) with trypsin-like serine proteinases. Biol Chem Hoppe Seyler. 1986 Sep;367(9):853–859. doi: 10.1515/bchm3.1986.367.2.853. [DOI] [PubMed] [Google Scholar]
  28. Yan S. C., Razzano P., Chao Y. B., Walls J. D., Berg D. T., McClure D. B., Grinnell B. W. Characterization and novel purification of recombinant human protein C from three mammalian cell lines. Biotechnology (N Y) 1990 Jul;8(7):655–661. doi: 10.1038/nbt0790-655. [DOI] [PubMed] [Google Scholar]
  29. de Roux N., Chadeuf G., Molho-Sabatier P., Plouin P. F., Aiach M. Clinical and biochemical characterization of antithrombin III Franconville, a variant with Pro 41 Leu mutation. Br J Haematol. 1990 Jun;75(2):222–227. doi: 10.1111/j.1365-2141.1990.tb02653.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES