Download PDF
Thromb Haemost 1998; 80(04): 585-587
DOI: 10.1055/s-0037-1615425
Rapid Communication
Schattauer GmbH

Activity of Secreted Kunitz Domain 1 Variants of Tissue Factor Pathway Inhibitor

Kirk Johnson   *
,
Isabel Zaror   *
,
Diane Bauer
1   From the Chiron Technologies, Chiron Corporation, Emeryville, CA, USA
,
Yoon Choi
1   From the Chiron Technologies, Chiron Corporation, Emeryville, CA, USA
,
Abla Creasey
1   From the Chiron Technologies, Chiron Corporation, Emeryville, CA, USA
,
Michael Innis
1   From the Chiron Technologies, Chiron Corporation, Emeryville, CA, USA
› Author Affiliations
Further Information

Publication History

Received 30 June 1997

Accepted after resubmission 17 June 1998

Publication Date:
08 December 2017 (online)

    Permissions and Reprints

Summary

Tissue factor pathway inhibitors (TFPI and TFPI-2) are Kunitz domain-type serine protease inhibitors which inhibit factor VIIa/tissue factor (VIIa/TF) complexes in a factor Xa-dependent manner. The VIIa/TF and Xa inhibitory activity has been localized to the first two Kunitz domains, respectively. Unlike TFPI, TFPI-2 has been reported to exhibit significant Xa-independent VIIa/TF inhibitory activity, perhaps due to an arginine at the P1 residue in the first Kunitz domain of TFPI-2 as opposed to a lysine at the comparable residue in TFPI. Two domain TFPI variants, differing in the first Kunitz domain but containing the second Kunitz domain of TFPI, were constructed and secreted by Saccharomyces cerevisiae in order to test the possibility that a TFPI first Kunitz domain with a P1 lysine to arginine change or a hybrid containing the TFPI-2 first Kunitz domain may represent more potent VIIa/TF inhibitors. When yeast supernatants were analyzed for specific activity in the Xa-dependent inhibition of VIIa/TF, neither variant was as active as the truncated TFPI.

* Authors contributed equally


 

  • References

  • 1 Davie EW, Fujikawa K, Kisiel W. The coagulation cascade: Initiation, maintenance, and regulation.. Biochemistry 1991; 30: 10363-70.
    CrossrefPubMedGoogle Scholar
  • 2 Broze Jr. GJ. The role of tissue factor pathway inhibitor in a revised coagulation cascade.. Semin Hematol 1992; 29: 159-69.
    PubMedGoogle Scholar
  • 3 Wun TC, Kretzmer KK, Girard TJ, Miletich JP, Broze Jr. GJ. Cloning and characterization of a cDNA coding for the lipoprotein-associated coagulation inhibitor shows that it consists of three tandem Kunitz-type inhibitory domains.. J Biol Chem 1988; 263: 6001-4.
    PubMedGoogle Scholar
  • 4 Girard TJ, Warren LA, Novotny WF, Likert KM, Brown SG, Miletich JP. et al. Functional significance of the Kunitz-type inhibitory domains of lipoprotein-associated coagulation inhibitor.. Nature 1989; 338: 518-20.
    CrossrefPubMedGoogle Scholar
  • 5 Enjyoji K, Miyata T, Kamikubo Y, Kato H. Effect of heparin on the inhibition of factor Xa by tissue factor pathway inhibitor: a segment, Gly212-Phe243, of the third Kunitz domain is a heparin-binding site.. Biochemistry 1995; 34: 17 5725-35.
    PubMedGoogle Scholar
  • 6 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI).. Thromb Res 1988; 50: 803-5.
    CrossrefPubMedGoogle Scholar
  • 7 Petersen JG, Meyn G, Rasmussen JS, Petersen J, Bjorn SE, Jonassen I, Christiansen L, Nordfang O. Characterization of human tissue factor pathway inhibitor variants expressed in Saccharomyces cerevisiae.. J Biol Chem 1993; 268: 13344-51.
    PubMedGoogle Scholar
  • 8 Wesselschmidt R, Likert K, Girard T, Wun TC, Broze Jr. GJ. Tissue factor pathway inhibitor: the carboxy-terminus is required for optimal inhibition of factor Xa.. Blood 1992; 79: 2004-10.
    PubMedGoogle Scholar
  • 9 Sprecher CA, Kisiel W, Mathewes S, Foster DC. Evidence that a second human tissue factor pathway inhibitor (TFPI-2) and human placental protein 5 are equivalent.. Proc Natl Acad Sci USA 1994; 91: 3353-7.
    CrossrefPubMedGoogle Scholar
  • 10 Dennis MS, Lazarus RA. Kunitz domain inhibitors of tissue factor-factor VIIa. I. Potent inhibitors selected from libraries by phage display.. J Biol Chem 1994; 269: 22129-36.
    PubMedGoogle Scholar
  • 11 Maniatis T, Fritsch EF, Sambrook J. Ed. Molecular Cloning: A Laboratory Manual.. 1982 Cold Spring Harbor Laboratory,. New York.:
    PubMedGoogle Scholar
  • 12 Brake AJ. Alpha-factor leader-directed secretion of heterologous proteins from yeast. Meth.. Enzymol 1990; 185: 408-21.
    PubMedGoogle Scholar
  • 13 Barr PJ, Cousens LS, Lee-Ng CT, Medina-Selby A, Masiarz FR, Hallewell RA, Chamberlain S, Bradley J, Steimer KS, Esch F, Baird AJ. Expression and processing of biologically active fibroblast growth factors in the yeast Saccharomyces cerevisiae.. J Biol Chem 1988; 263: 16471-8.
    PubMedGoogle Scholar
  • 14 Cousens LS, Shuster JR, Gallegos C, Ku L, Stempien MM, Urdea MS, Sanchez-Pescador R, Taylor A, Tekamp-Olson P. High level expression of proinsulin in the yeast, Saccharomyces cerevisiae.. Gene 1987; 61: 265-75.
    CrossrefPubMedGoogle Scholar
  • 15 Julius D, Brake A, Blair L, Kunisawa R, Thorner J. Isolation of the putative structural gene for the lysine-arginine-cleaving endopeptidase required for processing of yeast prepro-alpha-factor.. Cell 1984; 37: 1075-89.
    CrossrefPubMedGoogle Scholar
  • 16 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 1970; 227: 680-5.
    CrossrefPubMedGoogle Scholar
  • 17 Girard TJ, Broze Jr. GJ. Tissue factor pathway inhibitor meth.. Enzymol 1993; 222: 195-209.
    PubMedGoogle Scholar
  • 18 Dennis MS, Lazarus RA. Kunitz domain inhibitors of tissue factor-factor VIIa. II. Potent and specific inhibitors by competitive phage selection.. J Biol Chem 1994; 269: 22137-44.
    PubMedGoogle Scholar
  • 19 Petersen LC, Bjorn SE, Olsen OH, Nordfang O, Norris F, Norris K. Inhibitory properties of separate recombinant Kunitz-type-protease-inhibitor domains from tissue-factor-pathway inhibitor.. Eur J Biochem 1996; 235: 310-16.
    CrossrefPubMedGoogle Scholar