Subscribe to RSS
DOI: 10.1055/s-0037-1615425
Activity of Secreted Kunitz Domain 1 Variants of Tissue Factor Pathway Inhibitor
Publication History
Received
30 June 1997
Accepted after resubmission
17 June 1998
Publication Date:
08 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199804/lookinside/thumbnails/10-1055-s-0037-1615425-1.jpg)
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.
- 2 Broze Jr. GJ. The role of tissue factor pathway inhibitor in a revised coagulation cascade.. Semin Hematol 1992; 29: 159-69.
- 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.
- 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.
- 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.
- 6 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI).. Thromb Res 1988; 50: 803-5.
- 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.
- 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.
- 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.
- 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.
- 11 Maniatis T, Fritsch EF, Sambrook J. Ed. Molecular Cloning: A Laboratory Manual.. 1982 Cold Spring Harbor Laboratory,. New York.:
- 12 Brake AJ. Alpha-factor leader-directed secretion of heterologous proteins from yeast. Meth.. Enzymol 1990; 185: 408-21.
- 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.
- 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.
- 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.
- 16 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 1970; 227: 680-5.
- 17 Girard TJ, Broze Jr. GJ. Tissue factor pathway inhibitor meth.. Enzymol 1993; 222: 195-209.
- 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.
- 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.