Thromb Haemost 1989; 62(03): 902-905
DOI: 10.1055/s-0038-1651025
Original Article
Schattauer GmbH Stuttgart

Molecular Forms of Human Protein C: Comparison and Distribution in Human Adult Plasma

Brian S Greffe
1   The Pediatric Hematology, Department of Pediatrics, Denver, Colorado, USA
,
Marilyn J Manco-Johnson
1   The Pediatric Hematology, Department of Pediatrics, Denver, Colorado, USA
,
Richard A Marlar
1   The Pediatric Hematology, Department of Pediatrics, Denver, Colorado, USA
2   The Department of Biochemistry, University of Colorado Health Sciences Center, Denver, Colorado, USA
3   The Laboratory Services, Denver Veterans Administration Medical Center, Denver, Colorado, USA
› Author Affiliations
Further Information

Publication History

Received 15 February 1989

Accepted after revision 28 July 1989

Publication Date:
30 June 2018 (online)

Summary

Protein C (PC) is a vitamin K-dependent protein which functions as both an anticoagulant and profibrinolytic. It is synthesized as a single chain protein (SC-PC) and post-transla-tionally modified into a two chain form (2C-PC). Two chain PC consists of a light chain (LC) and a heavy chain (HC). The present study was undertaken to determine the composition of the molecular forms of PC in plasma. PC was immunoprecipitated, subjected to SDS-PAGE and Western blotting. The blots were scanned by densitometry to determine the distribution of the various forms. The percentage of SC-PC and 2C-PC was found to be 10% and 90% respectively. This is in agreement with previous work. SC-PC and the heavy chain of 2C-PC consisted of three molecular forms (“alpha”, “beta”, and “gamma”). The “alpha” form of HC is the standard 2C form with a MW of 40 Kd. The “beta” form of HC has also been described and has MW which is 4 Kd less than the “alpha” form. The “gamma” species of the SC and 2C-PC has not been previously described. However, its 3 Kd difference from the “beta” form could be due to modification of the “beta” species or to a separate modification of the alpha-HC. The LC of PC was shown to exist in two forms (termed form 1 and form 2). The difference between these two forms is unknown. The molecular forms of PC are most likely due to a post-translational modification (either loss of a carbohydrate or a peptide) rather than from plasma derived degradation.

 
  • References

  • 1 Clouse LH, Comp PC. The regulation of hemostasis: The protein C system. NEJM 1986; 314: 1298-1304
  • 2 Esmon CT. The regulation off natural anticoagulant pathways. Science 1987; 235: 1348-1352
  • 3 Fair DS, Marlar RA. Biosynthesis and secretion of factor VII, protein C, protein S, and the protein C in inhibitor from human hepatoma cell line. Blood 1986; 67: 64-70
  • 4 Plutzky J, Hoskins J, Long GL, Crabtree GR. Evolution and organization of the human protein C gene. Proc Natl Acad Sci USA 1986; 83: 546-550
  • 5 Kisiel W. Human plasma protein C: Isolation, characterization, and mechanism of activation by thrombin. J Clin Invest 1979; 64: 761-769
  • 6 Marlar RA, Kleiss AJ, Griffin JH. Mechanism of action of human activated protein C, a thrombin dependent anticoagulation enzyme. Blood 1982; 59: 1067-1072
  • 7 Stenflo J. Structure and function of protein C. Semin Thromb Haemostas 1984; 10: 109-121
  • 8 Graves CB, Munns TW, Willingham AK. Rat factor-X is synthesized as a single chain precursor inducible by prothrombin fragments. J Biol Chem 1982; 257: 13108-13113
  • 9 Kisiel W, Davie EW. protein C. Meth Enzymol 1981; 80: 320-332
  • 10 Heeb MJ, Schwarz P, White T, Lammle B, Berrettini M, Griffin JH. Immunoblotting studies of the molecular forms of protein C in plasma. Thromb Res 1988; 52: 33-43
  • 11 Foster DC, Yoshitake S, Davie EW. The nucleotide sequence of the gene for human protein C. Proc Natl Acad Sci USA 1985; 82: 4673-4677
  • 12 Long GL. Structure and evolution of the human genes encoding protein C and coagulation factors VII, IX, and X, Cold Spring Harbor Symp. Quantitative Biol 1986; 51: 525-529
  • 13 Furie B, Furie BC. The molecular basis of blood coagulation. Cell 1988; 53: 505-518
  • 14 Marlar RA. Plasma single chain protein C is functionally similar to the two chain form of plasma protein C. Thromb Haemostas 1985; 54: 1275
  • 15 Miletich JP, Leykam JF, Broze GJ. Detection of single chain protein C in human plasma. Blood 1983; (Suppl 1) 62: 306a
  • 16 Hathaway WE, Bonnar J. Technical aspects of hemostatic measurements. In: Hemostatic Disorders of the Pregnant Woman and Newborn Infant. Elsevier; New York: 1987: 29
  • 17 Marlar RA, Madden RM, Oppenheimer C, Wydro R. Functional studies of recombinant protein C. Fibrinolysis 1988; (Suppl 1) 2: 154 (Abstr).
  • 18 Oppenheimer C, Wydro R. Cellular processing of vitamin K dependent proteins. In: Current Advances in Vitamin K Research Suttie JW. (ed). Elsevier; New York: 1988: 165-171
  • 19 Laurell M, Ikeda K, Stenflo J. Characterization of monoclonal antibodies against human protein C specific for the calcium ion-induced conformation or for the acivation peptide region. FEBS Lett 1985; 191: 75-81
  • 20 Manco-Johnson MJ, Marlar RA, Jacobson LJ, Hays T, Warady BA. Severe protein C deficiency in newborn infants. J Pediatr 1988; 113: 359-363
  • 21 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
  • 22 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
  • 23 Marlar RA, Endres Brooks, Miller C. Serial studies of protein C and its plasma inhibitor in patients with disseminated intravascular coagulation. Blood 1985; 66: 59-63
  • 24 Miletich JP, Broze GJ. Human p-Protein C is not glycosylated at the fourth N-linked site. Blood 1988; 72: 371a
  • 25 Fujikawa K, Titani K, Davie EW. Activation of bovine factor X (Stuart factor): Conversion of factor Xα to factor Xβ. Proc Natl Acad Sci USA 1975; 72: 3359-3363
  • 26 Marlar RA, Seegers WH. Formation and properties of thrombin generated factor IX inhibitor. Thromb Res 1979; 14: 095-105