Abstract
cDNA coding for protein C has been cloned from a bovine liver library in plasmid vector pBR322 and its sequence has been determined. Two overlapping clones code for the entire light and heavy chains of the mature protein, as well as a previously unreported connecting dipeptide (Lys-Arg) and a 39-amino acid leader peptide region. Identification and characterization of the clones establishes the liver as a site of protein C biosynthesis. A contiguous coding region reveals that a one-chain precursor protein is made that upon limited proteolysis yields both the mature light and heavy chains. The codon for aspartic acid is found at light chain amino acid position 71, showing that the beta-hydroxyaspartic acid that exists in this position of the mature protein is the result of post-translational modification of an aspartic acid residue. Amino acid sequence homology in the amino-terminal region of the light chain with other vitamin K-dependent coagulation factors is continued into the leader peptide region.
Full text
PDF



Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- 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]
- 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]
- 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]
- Esmon C. T. Protein-C: biochemistry, physiology, and clinical implications. Blood. 1983 Dec;62(6):1155–1158. [PubMed] [Google Scholar]
- Esmon C. T., Stenflo J., Suttie J. W. A new vitamin K-dependent protein. A phospholipid-binding zymogen of a serine esterase. J Biol Chem. 1976 May 25;251(10):3052–3056. [PubMed] [Google Scholar]
- 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]
- Fernlund P., Stenflo J. Amino acid sequence of the light chain of bovine protein C. J Biol Chem. 1982 Oct 25;257(20):12170–12179. [PubMed] [Google Scholar]
- Fernlund P., Stenflo J., Tufvesson A. Bovine protein C: amino acid sequence of the light chain. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5889–5892. doi: 10.1073/pnas.75.12.5889. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fung M. R., Campbell R. M., MacGillivray R. T. Blood coagulation factor X mRNA encodes a single polypeptide chain containing a prepro leader sequence. Nucleic Acids Res. 1984 Jun 11;12(11):4481–4492. doi: 10.1093/nar/12.11.4481. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Kisiel W., Canfield W. M., Ericsson L. H., Davie E. W. Anticoagulant properties of bovine plasma protein C following activation by thrombin. Biochemistry. 1977 Dec 27;16(26):5824–5831. doi: 10.1021/bi00645a029. [DOI] [PubMed] [Google Scholar]
- Kisiel W., Ericsson L. H., Davie E. W. Proteolytic activation of protein C from bovine plasma. Biochemistry. 1976 Nov 2;15(22):4893–4900. doi: 10.1021/bi00667a022. [DOI] [PubMed] [Google Scholar]
- Kisiel W. Human plasma protein C: isolation, characterization, and mechanism of activation by alpha-thrombin. J Clin Invest. 1979 Sep;64(3):761–769. doi: 10.1172/JCI109521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Land H., Grez M., Hauser H., Lindenmaier W., Schütz G. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency. Nucleic Acids Res. 1981 May 25;9(10):2251–2266. doi: 10.1093/nar/9.10.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacGillivray R. T., Davie E. W. Characterization of bovine prothrombin mRNA and its translation product. Biochemistry. 1984 Apr 10;23(8):1626–1634. doi: 10.1021/bi00303a007. [DOI] [PubMed] [Google Scholar]
- MacGillivray R. T., Degen S. J., Chandra T., Woo S. L., Davie E. W. Cloning and analysis of a cDNA coding for bovine prothrombin. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5153–5157. doi: 10.1073/pnas.77.9.5153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Olson R. E., Suttie J. W. Vitamin K and gamma-carboxyglutamate biosynthesis. Vitam Horm. 1977;35:59–108. doi: 10.1016/s0083-6729(08)60521-x. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Sanger F., Coulson A. R. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. doi: 10.1016/0014-5793(78)80145-8. [DOI] [PubMed] [Google Scholar]
- Seegers W. H., Marlar R. A., Walz D. A. Anticoagulant effects of autoprothrombin II-A and prothrombin fragment 1. Thromb Res. 1978 Aug;13(2):233–243. doi: 10.1016/0049-3848(78)90011-7. [DOI] [PubMed] [Google Scholar]
- Seegers W. H., Novoa E., Henry R. L., Hassouna H. I. Relationship of "new" vitamin K-dependent Protein C and "old" autoprothrombin II-a. Thromb Res. 1976 May;8(5):543–552. doi: 10.1016/0049-3848(76)90236-x. [DOI] [PubMed] [Google Scholar]
- Smith G. E., Summers M. D. The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980 Nov 15;109(1):123–129. doi: 10.1016/0003-2697(80)90019-6. [DOI] [PubMed] [Google Scholar]
- Stenflo J. A new vitamin K-dependent protein. Purification from bovine plasma and preliminary characterization. J Biol Chem. 1976 Jan 25;251(2):355–363. [PubMed] [Google Scholar]
- Stenflo J., Fernlund P. Amino acid sequence of the heavy chain of bovine protein C. J Biol Chem. 1982 Oct 25;257(20):12180–12190. [PubMed] [Google Scholar]
- Vehar G. A., Davie E. W. Preparation and properties of bovine factor VIII (antihemophilic factor). Biochemistry. 1980 Feb 5;19(3):401–410. doi: 10.1021/bi00544a001. [DOI] [PubMed] [Google Scholar]
- Walker F. J., Sexton P. W., Esmon C. T. The inhibition of blood coagulation by activated Protein C through the selective inactivation of activated Factor V. Biochim Biophys Acta. 1979 Dec 7;571(2):333–342. doi: 10.1016/0005-2744(79)90103-7. [DOI] [PubMed] [Google Scholar]