Skip to main content
NCBI 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
. 1987 Jan;84(2):349–353. doi: 10.1073/pnas.84.2.349

Cloning and characterization of human liver cDNA encoding a protein S precursor.

J Hoskins, D K Norman, R J Beckmann, G L Long
PMCID: PMC304204  PMID: 3467362

Abstract

Human liver cDNA encoding a protein S precursor was isolated from two cDNA libraries by two different techniques. Based upon the frequency of positive clones, the abundance of mRNA for protein S is approximately 0.01%. Blot hybridization of electrophoretically fractionated poly(A)+ RNA revealed a major mRNA approximately 4 kilobases long and two minor forms of approximately 3.1 and approximately equal to 2.6 kilobases. One of the cDNA clones contains a segment encoding a 676 amino acid protein S precursor, as well as 108 and 1132 nucleotides of 5' and 3' noncoding sequence, respectively, plus a poly(A) region at the 3' end. The cDNAs are adenosine plus thymidine-rich (60%) except for the 5' noncoding region, where 78% of the nucleotides are guanosine or cytosine. The protein precursor consists of a 41 amino acid "leader" peptide followed by 635 amino acids corresponding to mature protein S. Comparison of the mature protein region with homologous vitamin K-dependent plasma proteins shows that it is composed of the following domains: an amino-terminal gamma-carboxyglutamic acid-rich region of 37 amino acids; a 36 amino acid linker region rich in hydroxy amino acids; four epidermal growth factor-like segments, each approximately 45 amino acids long; and a 387 amino acid carboxyl-terminal domain of unrecognized structure and unknown function.

Full text

PDF
349

Images in this article

352Image
on p.352

Selected References

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

  1. Anson D. S., Choo K. H., Rees D. J., Giannelli F., Gould K., Huddleston J. A., Brownlee G. G. The gene structure of human anti-haemophilic factor IX. EMBO J. 1984 May;3(5):1053–1060. doi: 10.1002/j.1460-2075.1984.tb01926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beckmann R. J., Schmidt R. J., Santerre R. F., Plutzky J., Crabtree G. R., Long G. L. The structure 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 Acids Res. 1985 Jul 25;13(14):5233–5247. doi: 10.1093/nar/13.14.5233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bertina R. M. Hereditary protein S deficiency. Haemostasis. 1985;15(4):241–246. doi: 10.1159/000215155. [DOI] [PubMed] [Google Scholar]
  4. Bertina R. M., van Wijngaarden A., Reinalda-Poot J., Poort S. R., Bom V. J. Determination of plasma protein S--the protein cofactor of activated protein C. Thromb Haemost. 1985 Apr 22;53(2):268–272. [PubMed] [Google Scholar]
  5. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Breathnach R., Benoist C., O'Hare K., Gannon F., Chambon P. Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4853–4857. doi: 10.1073/pnas.75.10.4853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Broekmans A. W., Bertina R. M., Reinalda-Poot J., Engesser L., Muller H. P., Leeuw J. A., Michiels J. J., Brommer E. J., Briët E. Hereditary protein S deficiency and venous thrombo-embolism. A study in three Dutch families. Thromb Haemost. 1985 Apr 22;53(2):273–277. [PubMed] [Google Scholar]
  8. Bányai L., Váradi A., Patthy L. Common evolutionary origin of the fibrin-binding structures of fibronectin and tissue-type plasminogen activator. FEBS Lett. 1983 Oct 31;163(1):37–41. doi: 10.1016/0014-5793(83)81157-0. [DOI] [PubMed] [Google Scholar]
  9. Comp P. C., Esmon C. T. Recurrent venous thromboembolism in patients with a partial deficiency of protein S. N Engl J Med. 1984 Dec 13;311(24):1525–1528. doi: 10.1056/NEJM198412133112401. [DOI] [PubMed] [Google Scholar]
  10. Comp P. C., Nixon R. R., Cooper M. R., Esmon C. T. Familial protein S deficiency is associated with recurrent thrombosis. J Clin Invest. 1984 Dec;74(6):2082–2088. doi: 10.1172/JCI111632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  12. Dahlbäck B., Hildebrand B. Degradation of human complement component C4b in the presence of the C4b-binding protein-protein S complex. Biochem J. 1983 Mar 1;209(3):857–863. doi: 10.1042/bj2090857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dahlbäck B. Interaction between vitamin K-dependent protein S and the complement protein, C4b-binding protein. A link between coagulation and the complement system. Semin Thromb Hemost. 1984 Apr;10(2):139–148. doi: 10.1055/s-2007-1004416. [DOI] [PubMed] [Google Scholar]
  14. Dahlbäck B. Purification of human C4b-binding protein and formation of its complex with vitamin K-dependent protein S. Biochem J. 1983 Mar 1;209(3):847–856. doi: 10.1042/bj2090847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dahlbäck B. Purification of human vitamin K-dependent protein S and its limited proteolysis by thrombin. Biochem J. 1983 Mar 1;209(3):837–846. doi: 10.1042/bj2090837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Degen S. J., MacGillivray R. T., Davie E. W. Characterization of the complementary deoxyribonucleic acid and gene coding for human prothrombin. Biochemistry. 1983 Apr 26;22(9):2087–2097. doi: 10.1021/bi00278a008. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Drakenberg T., Fernlund P., Roepstorff P., Stenflo J. beta-Hydroxyaspartic acid in vitamin K-dependent protein C. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1802–1806. doi: 10.1073/pnas.80.7.1802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ebina Y., Ellis L., Jarnagin K., Edery M., Graf L., Clauser E., Ou J. H., Masiarz F., Kan Y. W., Goldfine I. D. The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell. 1985 Apr;40(4):747–758. doi: 10.1016/0092-8674(85)90334-4. [DOI] [PubMed] [Google Scholar]
  20. Fernlund P., Stenflo J. Beta-hydroxyaspartic acid in vitamin K-dependent proteins. J Biol Chem. 1983 Oct 25;258(20):12509–12512. [PubMed] [Google Scholar]
  21. Fung M. R., Hay C. W., MacGillivray R. T. Characterization of an almost full-length cDNA coding for human blood coagulation factor X. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3591–3595. doi: 10.1073/pnas.82.11.3591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Gilbert W. Why genes in pieces? Nature. 1978 Feb 9;271(5645):501–501. doi: 10.1038/271501a0. [DOI] [PubMed] [Google Scholar]
  23. Griffin J. H. Clinical studies of protein C. Semin Thromb Hemost. 1984 Apr;10(2):162–166. doi: 10.1055/s-2007-1004419. [DOI] [PubMed] [Google Scholar]
  24. Hagen F. S., Gray C. L., O'Hara P., Grant F. J., Saari G. C., Woodbury R. G., Hart C. E., Insley M., Kisiel W., Kurachi K. Characterization of a cDNA coding for human factor VII. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2412–2416. doi: 10.1073/pnas.83.8.2412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jackson R. C., Blobel G. Post-translational processing of full-length presecretory proteins with canine pancreatic signal peptidase. Ann N Y Acad Sci. 1980;343:391–404. doi: 10.1111/j.1749-6632.1980.tb47268.x. [DOI] [PubMed] [Google Scholar]
  26. Katz L., Kingsbury D. T., Helinski D. R. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J Bacteriol. 1973 May;114(2):577–591. doi: 10.1128/jb.114.2.577-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lai E. C., Riser M. E., O'Malley B. W. Regulated expression of the chicken ovalbumin gene in a human estrogen-responsive cell line. J Biol Chem. 1983 Oct 25;258(20):12693–12701. [PubMed] [Google Scholar]
  28. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  29. Lundwall A., Dackowski W., Cohen E., Shaffer M., Mahr A., Dahlbäck B., Stenflo J., Wydro R. Isolation and sequence of the cDNA for human protein S, a regulator of blood coagulation. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6716–6720. doi: 10.1073/pnas.83.18.6716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  31. McMullen B. A., Fujikawa K. Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). J Biol Chem. 1985 May 10;260(9):5328–5341. [PubMed] [Google Scholar]
  32. Nelsestuen G. L., Kisiel W., Di Scipio R. G. Interaction of vitamin K dependent proteins with membranes. Biochemistry. 1978 May 30;17(11):2134–2138. doi: 10.1021/bi00604a017. [DOI] [PubMed] [Google Scholar]
  33. Ovchinnikov Y. A., Monastyrskaya G. S., Gubanov V. V., Guryev S. O., Chertov OYu, Modyanov N. N., Grinkevich V. A., Makarova I. A., Marchenko T. V., Polovnikova I. N. The primary structure of Escherichia coli RNA polymerase. Nucleotide sequence of the rpoB gene and amino-acid sequence of the beta-subunit. Eur J Biochem. 1981 Jun 1;116(3):621–629. doi: 10.1111/j.1432-1033.1981.tb05381.x. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Perlman D., Halvorson H. O. A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J Mol Biol. 1983 Jun 25;167(2):391–409. doi: 10.1016/s0022-2836(83)80341-6. [DOI] [PubMed] [Google Scholar]
  36. Schwarz H. P., Fischer M., Hopmeier P., Batard M. A., Griffin J. H. Plasma protein S deficiency in familial thrombotic disease. Blood. 1984 Dec;64(6):1297–1300. [PubMed] [Google Scholar]
  37. Stenflo J. Structure and function of protein C. Semin Thromb Hemost. 1984 Apr;10(2):109–121. doi: 10.1055/s-2007-1004413. [DOI] [PubMed] [Google Scholar]
  38. Suzuki K., Nishioka J., Hashimoto S. Regulation of activated protein C by thrombin-modified protein S. J Biochem. 1983 Sep;94(3):699–705. doi: 10.1093/oxfordjournals.jbchem.a134409. [DOI] [PubMed] [Google Scholar]
  39. Ullrich A., Bell J. R., Chen E. Y., Herrera R., Petruzzelli L. M., Dull T. J., Gray A., Coussens L., Liao Y. C., Tsubokawa M. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. 1985 Feb 28-Mar 6Nature. 313(6005):756–761. doi: 10.1038/313756a0. [DOI] [PubMed] [Google Scholar]
  40. Walker F. J. Regulation of activated protein C by a new protein. A possible function for bovine protein S. J Biol Chem. 1980 Jun 25;255(12):5521–5524. [PubMed] [Google Scholar]
  41. Walker F. J. Regulation of activated protein C by protein S. The role of phospholipid in factor Va inactivation. J Biol Chem. 1981 Nov 10;256(21):11128–11131. [PubMed] [Google Scholar]
  42. Walker F. J. Regulation of bovine activated protein C by protein S: the role of the cofactor protein in species specificity. Thromb Res. 1981 May 1;22(3):321–327. doi: 10.1016/0049-3848(81)90125-0. [DOI] [PubMed] [Google Scholar]
  43. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [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