Human endothelial cells synthesize protein Z, but not the protein Z dependent inhibitor

 

 Buy Article Permissions and Reprints

Summary

Protein Z (PZ) is a vitamin K-dependent protein isolated from human plasma, and acts as a cofactor fora serpin, called protein Z-dependent protease inhibitor (ZPI).A prothrombotic phenotype has been reported in PZ deficient mice, and PZ deficiencies have been observed in patients with arterial thrombotic events. PZ was immunologically detected in the endothelium of atherosclerotic arteries, suggesting that endothelial cells could be involved in the production of PZ.In this study we analyzed the synthesis and release of PZ and ZPI by human umbilical vein endothelial cells (HUVEC), representative of the macrovasculature, and by HMEC-1, a microvascular endothelial cell line. PZ was quantified bya specific ELISA in the supernatant and in the lysates of both cellular types. Western blotting of the supernatants showed the presence of a band of 62 kDa,identical to PZ synthesized by the hepatoma cell line HepG2. mRNA of PZ was also detected in each cellular type. PZ biosynthesis was unaffected by inflammatory cytokines in HUVEC, whereas a slight decrease of mRNA and PZ antigen (53.5 ± 14.5% of protein synthesis as compared to the control, p < 0.01) and a modest increase (126± 8.5% as compared to the control, p< 0.05) were induced respectively byTumor Necrosis Factor (TNF)-alpha (25 ng/ml) and oncostatin M (5 ng/ml) in HMEC-1. Immunological studies showed the presence of PZ near the nucleus and a possible expression of PZ at the membrane.In addition, PZ was present in the endothelial cells of both normal arterial and venous vessel sections. In contrast, neither ZPI nor its mRNA was detected in endothelial cells.

• References

• 1 Broze Jr GJ, Miletich JP. Human protein Z. J Clin Invest 1984; 73: 933-8.
  CrossrefPubMedGoogle Scholar
• 2 Ichinose A, Takeya H, Espling E. et al. Amino acid sequence of human protein Z, a vitamin K-dependent plasma glycoprotein. Biochem Biophys Res Commun 1990; 172: 1139-44.
  CrossrefPubMedGoogle Scholar
• 3 Walker FJ. Regulation of coagulation by activated proteinC by protein S. The role of phospholipid in factor Va inactivation. J Biol Chem 1981; 256: 11128-31.
  PubMedGoogle Scholar
• 4 Han X, Huang ZF, Fiehler R. et al. The protein Z-dependent protease inhibitor isa serpin. Biochemistry 1999; 38: 11073-8.
  CrossrefPubMedGoogle Scholar
• 5 Tabatabai A, Fiehler R, Broze Jr GJ. Protein Z circulates in plasma in a complex with protein Z-dependent protease inhibitor. Thromb Haemost 2001; 85: 655-60.
  Article in Thieme ConnectPubMedGoogle Scholar
• 6 Han X, Fiehler R, Broze Jr GJ. Isolation of a protein Z-dependent plasma protease inhibitor. Proc Natl Acad Sci USA 1998; 95: 9250-5.
  CrossrefPubMedGoogle Scholar
• 7 Yin Z-F, Huang ZF, Cui J. et al. Prothrombotic phenotype of protein Z deficiency. Proc Natl Acad Sci USA 2000; 97: 6734-8.
  CrossrefPubMedGoogle Scholar
• 8 Vasse M, Guegan-Massardier E, Borg J-Y. et al. High frequency of protein Z deficiency in patients with ischemic stroke. Lancet 2001; 357: 933-4.
  CrossrefPubMedGoogle Scholar
• 9 Heeb MJ, Paganini-Hill A, Griffin JH. et al. Low protein Z levels and risk of ischemic stroke: differences by diabetic status and gender. Blood Cells Mol Dis 2002; 29: 139-44.
  CrossrefPubMedGoogle Scholar
• 10 Fedi S, Sofi F, Brogi D. et al. Low protein Z plasma levels are independently associated with acute coronary syndromes. Thromb Haemost 2003; 90: 1173-8.
  Article in Thieme ConnectPubMedGoogle Scholar
• 11 Ayoub N, Esposito G, Barete S. et al. Protein Z deficiency in antiphospholipid-negative Sneddon's syndrome. Stroke 2004; 35: 1329-32.
  CrossrefPubMedGoogle Scholar
• 12 Al-Shanqeeti A, van Hylckama Vlieg A, Berntorp E. et al. Protein Z and protein Z-dependent protease inhibitor. Determinants of levels and risk of venous thrombosis. Thromb Haemost 2005; 93: 411-3.
  Article in Thieme ConnectPubMedGoogle Scholar
• 13 Gris JC, Quéré I, Dechaud H. et al. High frequency of protein Z deficiency in patients with unexplained early fetal loss. Blood 2001; 99: 2606-8.
  PubMedGoogle Scholar
• 14 Kemkes-Matthes B, Matthes KJ. Protein Z, a new haemostatic factor, in liver diseases. Haemostasis 1995; 25: 312-6.
  PubMedGoogle Scholar
• 15 Greten J, Kreis I, Liliensiek B. et al. Localisation of protein Z in vascular lesions of patients with atherosclerosis. Vasa 1998; 27: 144-8.
  PubMedGoogle Scholar
• 16 Vasse M, Pourtau J, Trochon V. et al. Oncostatin M induces angiogenesis in vitro and in vivo. Arterioscler Thromb Vasc Biol 1999; 19: 1835-42.
  CrossrefPubMedGoogle Scholar
• 17 Ades EW, Candal FJ, Swerlick RA. et al. HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J Invest Dermatol 1992; 99: 683-90.
  CrossrefPubMedGoogle Scholar
• 18 Vasse M, Denoyelle C, Legrand E. et al. Weak regulation of protein Z biosynthesis by inflammatory cytokines. Thromb Haemost 2002; 87: 350-1.
  Article in Thieme ConnectPubMedGoogle Scholar
• 19 Miletich JP, Broze Jr GJ. Human plasma protein Z antigen: range in normal subjects and effect of warfarin therapy. Blood 1987; 69: 1580-6.
  PubMedGoogle Scholar
• 20 Stern D, Brett J, Harris K. et al. Participation of endothelial cells in the protein C-proteinS anticoagulant pathway: the synthesis and release of protein S. J Cell Biol 1986; 102: 1971-8.
  CrossrefPubMedGoogle Scholar
• 21 Fair DS, Marlar RA, Levin EG. Human endothelial cells synthesize protein S. Blood 1986; 67: 1168-71.
  PubMedGoogle Scholar
• 22 Bajaj MS, Kuppuswamy MN, Saito H. et al. Cultured normal human hepatocytes do not synthesize lipoprotein-associated coagulation inhibitor: evidence that endothelium is the principal site of its synthesis. Proc Natl Acad Sci USA 1990; 87: 8869-73.
  CrossrefPubMedGoogle Scholar
• 23 Tanabe S, Sugo T, Matsude M. Synthesis of protein C in human umbilical vein endothelial cells. J Biochem 1991; 109: 924-8.
  CrossrefPubMedGoogle Scholar
• 24 Bertina R, Van Wijngaarden A, Reinalda-Poot J. et al. Determination of plasma PS – the protein cofactor of activated protein C. Thromb Haemost 1985; 53: 268-72.
  Article in Thieme ConnectPubMedGoogle Scholar
• 25 Soff GA, Jackman RW, Rosenberg RD. Expression of thrombomodulin by smooth muscle cells in culture: different effects of tumor necrosis factor and cyclic adenosine monophosphate on thrombomodulin expression by endothelial cells and smooth muscle cells in culture. Blood 1991; 77: 515-8.
  PubMedGoogle Scholar
• 26 Ross R. The pathogenesis of atherosclerosis:a perspective for the 1990s. Nature 1993; 362: 801-9.
  CrossrefPubMedGoogle Scholar
• 27 Modur V, Feldhaus MJ, Weyrich AS. et al. Oncostatin M is a proinflammatory mediator. In vivo effects correlate with endothelial cell expression of inflammatory cytokines and adhesion molecules. J Clin Invest 1997; 100: 158-68.
  CrossrefPubMedGoogle Scholar
• 28 Kobelt K, Biasiutti FD, Mattle HP. et al. Protein Z in ischaemic stroke. Brit J Haematol 2001; 114: 169-73.
  CrossrefPubMedGoogle Scholar
• 29 Staton J, Sayer M, Hankey GJ. et al. Protein Z polymorphisms, protein Z concentrations, and ischemic stroke. Stroke 2005; 36: 1123-7.
  CrossrefPubMedGoogle Scholar
• 30 Martinelli I, Razzari C, Biguzzi E. et al. Low levels of Protein Z and the risk of venous thromboembolism. J Thromb Haemost 2005; 12: 2817-9.
  PubMedGoogle Scholar