TFPIβ, a Second Product from the Mouse Tissue Factor Pathway Inhibitor (TFPI) Gene

Summary

Tissue factor pathway inhibitor (TFPI) contains three Kunitz domains separated by two connecting regions. We have cloned another naturally occurring TFPI gene product from a mouse lung cDNA library which we have called TFPI β. TFPIβ is derived from alternative splicing of the TFPI gene. Analysis of the cDNA shows that mouse TFPIβ protein is identical to TFPI from the N’-terminus through the second connecting region. However, mouse TFPIβ possesses neither a third Kunitz domain nor an Arg, Lys-rich C’-terminus but instead has a completely different C’-terminal (β-domain) sequence which is not homologous to any known protein. Northern blot analyses show that the tissues for mouse TFPIβ synthesis are heart and lung; in contrast, TFPI appears in Northern blots of heart and spleen. Both TFPIβ and TFPI messages first appear in 7-day-old mouse embryos, but only the TFPI mRNA persists until 17 days. Purified recombinant TFPIβ shows an apparent molecular weight of 38 kDa. Kinetic studies indicate that mouse TFPIβ is a slow-binding enzyme inhibitor for human factor Xa. In addition, heparin does not enhance the inhibition of factor Xa by mouse TFPIβ although it does accelerate factor Xa inhibition by TFPI.

 

• References

• 1 Wun T-Z, 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.
• 2 Kamei S, Kamikubo Y-i, Hamuro T, Fujimoto H, Ishihara M, Yonemura H, Miyamoto S, Funatsu A, Enjyoji K-i, Abumiya T, Miyata T, Kato H. Amino acid sequence and inhibitory activity of rhesus monkey tissue factor pathway inhibitor (TFPI): comparison with human TFPI. J Biochem 1994; 115: 708-14.
• 3 Girard TJ, Gailani D, Broze Jr. GJ. Complementary DNA sequencing of canine tissue factor pathway inhibitor reveals a unique nanomeric repetitive sequence between the second and third Kunitz domains. Biochem J 1994; 303: 923-8.
• 4 Wesselschmidt RL, Girard TJ, Broze Jr. GJ. cDNA sequence of rabbit lipoprotein-associated coagulation inhibitor. Nucleic Acids Res 1990; 18: 6440
• 5 Enjyoji K-i, Emi M, Mukai T, Kato H. cDNA Cloning and Expression of Rat Tissue Factor Pathway Inhibitor (TFPI). J Biochem 1992; 111: 681-7.
• 6 Chang J-Y, Monroe DM, Oliver JA, Liles DK, Roberts HR. Cloning, expression, and characterization of mouse tissue factor pathway inhibitor (TFPI). Thromb Haemost 1998; 79: 306-9.
• 7 Girard TJ, Warren LA, Novotny WF, Likert KM, Brown SG, Miletich JP, Broze Jr. GJ. Functional significance of the Kunitz-type inhibitory domains of lipoprotein-associated coagulation inhibitor. Nature 1989; 338: 518-20.
• 8 Wesselschmidt R, Likert K, Huang Z, MacPhail L, Broze Jr. GJ. Structural requirements for tissue factor pathway inhibitor interactions with factor Xa and heparin. Blood Coagul Fibrin 1993; 4: 661-9.
• 9 Enjyoji K-i, Miyata T, Kamikubo Y-i, 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: 5725-35.
• 10 Leff S, Rosenfeld M. Complex transcriptional units: diversity in gene expression by alternative RNA processing. Ann Rev Biochem 1986; 55: 1091-1117.
• 11 Maniatis T. Mechanisms of alternative pre-mRNA splicing. Science 1991; 251: 33-4.
• 12 Rio D. RNA binding proteins, splice site selection, and alternative pre-mRNA splicing. Gene Expression 1992; 2: 1-5.
• 13 Sharp P. Split genes and RNA splicing. Cell 1994; 77: 805-15.
• 14 Hynes RO. Fibronectins. Springer-Verlag; New York: 1990
• 15 Sanger F, Nicklen SL, Coulson AR. DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci U.S.A 1977; 74: 5463-7.
• 16 Wesselschmidt RL, Likert K, Girard T, Wun T-C, Broze Jr. GJ. Tissue factor pathway inhibitor: the carboxy-terminus is required for optimal inhibition of factor Xa. Blood 1992; 79: 2004-10.
• 17 Huang Z-F, Wun T-C, Broze Jr. GJ. Kinetics of factor Xa inhibition by tissue factor pathway inhibitor. J Biol Chem 1993; 268: 26950-5.
• 18 Engelman DM, Steitz TA, Goldman A. Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins. Ann Rev Biophys Biophys Chem 1986; 15: 321-53.
• 19 Girard TJ, Warren LA, Novonty WF, Bejcek BE, Miletich JP, Broze Jr. GJ. Identification of the 1.4 Kb and 4.0 Kb messages for the lipoprotein associated coagulation inhibitor and expression of the encoded protein. Thromb Res 1989; 55: 37-50.
• 20 Bajaj M, Kuppuswamy M, Satio H, Spitzer S, Bajaj S. Cultured normal human hepatocytes do not synthesize lipoprotein-associated coagulation inhibitor: evidence that endothelium is the principle site of its synthesis. Proc Natl Acad Sci USA 1990; 87: 8869-73.
• 21 Huang Z-F, Huguchi D, Lasky N, Broze Jr. GJ. Tissue factor pathway inhibitor gene disruption produces intrauterine lethality in mice. Blood 1997; 90: 944-51.
• 22 Kaufman MH. The Atlas of Mouse Development. Academic Press; New York: 1995
• 23 Kokawa T, Abumiya T, Kimura T, Harada-Shiba M, Koh H, Tsushima M, Yamamoto A, Kato H. Tissue factor pathway inhibitor activity in human plasma. Measurement of lipoprotein-associated and free forms in hyper-lipidemia. Arterioscler. Thromb Vasc Biol 1995; 15: 504-10.
• 24 Novotny WF, Girard TJ, Miletich J P, Broze Jr. GJ. Purification and characterization of the lipoprotein-associated coagulation inhibitor from human plasma. J Biol Chem 1989; 264: 18832-7.
• 25 Novotny WF, Girard TJ, Miletich JP, Broze Jr. GJ. Platelets secrete a coagulation inhibitor functionally and antigenically similar to the lipoprotein associated coagulation inhibitor. Blood 1988; 72: 2020-5.
• 26 Valentin S, Nordfang O, Bregengard C, Wildgoose P. Evidence that the C-terminus of tissue factor pathway inhibitor (TFPI) is essential for its in vitro and in vivo interaction with lipoproteins. Blood Coag Fibrinol 1993; 4: 713-20.
• 27 Novotny WF, Palmier M, Wun T-C, Broze Jr. GJ. Miletich JP. Purification and properties of heparin-releasable lipoprotein-associated coagulation inhibitor. Blood 1991; 78: 394-400.
• 28 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic pathway inhibitor (EPI). Thromb Res 1988; 50: 803-13.
• 29 Abumiya T, Enjyoji K-I, Kokawa T, Kamikubo Y-I, Kato H. An anti-tissue factor pathway inhibitor (TFPI) monoclonal antibody recognized the third Kunitz domain (K3) of free-form TFPI but not lipoprotein-associated forms in plasma. J. Biochem. 1995; 118: 178-82.
• 30 Gordon T, Castelli WP, Hjortland MC, Kannel WB, Dawber TR. High density lipoprotein as a protective factor against coronary heart disease. The Framingham study. Am J Med 1977; 62: 707-14.
• 31 Scanu AM. Lp(a) as a marker for coronary heart disease risk. Clin Cardiol 1991; 14: I-35-9.
• 32 Austin MA, Breslow JL, Hennekens CH, Buring JE, Willett WC. Krauss RM. Low-density lipoprotein subclass patterns and risk of myocardial infarction. JAMA 1988; 260: 1917-21.