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. 1998 Jan 15;101(2):321–326. doi: 10.1172/JCI1617

Generation and characterization of mice deficient in hepsin, a hepatic transmembrane serine protease.

Q Wu 1, D Yu 1, J Post 1, M Halks-Miller 1, J E Sadler 1, J Morser 1
PMCID: PMC508570  PMID: 9435303

Abstract

Hepsin is a type II transmembrane serine protease highly expressed on the surface of hepatocytes. The physiological function of hepsin is not known, although in vitro studies indicate that hepsin plays a role in the initiation of blood coagulation and in hepatocyte growth. To determine the functional importance of hepsin, we generated hepsin-deficient mice by homologous recombination. Homozygous hepsin-/- mice were viable and fertile, and grew normally. In functional assays including tail bleeding time, plasma clotting times, and tissue factor- or LPS-induced disseminated intravascular coagulation models, no significant difference was found between hepsin-/- and wild-type litter mates. Liver weight and serum concentrations of liver-derived proteins or enzymes were similar in hepsin-/- and wild-type mice. Interestingly, serum concentrations of bone-derived alkaline phosphatase were approximately twofold higher in hepsin-/- mice of both sexes when compared with wild-type litter mates. No obvious abnormalities were found in major organs in hepsin-/- mice in histological examinations. Our results indicate that hepsin is not essential for embryonic development and normal hemostasis. Hepsin-/- mice will help to evaluate the long-term effects of hepsin deficiency in these animals.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Appel L. F., Prout M., Abu-Shumays R., Hammonds A., Garbe J. C., Fristrom D., Fristrom J. The Drosophila Stubble-stubbloid gene encodes an apparent transmembrane serine protease required for epithelial morphogenesis. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):4937–4941. doi: 10.1073/pnas.90.11.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beaton A. H., Kiss I., Fristrom D., Fristrom J. W. Interaction of the Stubble-stubbloid locus and the Broad-complex of Drosophila melanogaster. Genetics. 1988 Oct;120(2):453–464. doi: 10.1093/genetics/120.2.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bond J. S. Plasma membrane proteases: introductory remarks. Biomed Biochim Acta. 1991;50(4-6):775–780. [PubMed] [Google Scholar]
  4. Broze G. J., Jr, Leykam J. E., Schwartz B. D., Miletich J. P. Purification of human brain tissue factor. J Biol Chem. 1985 Sep 15;260(20):10917–10920. [PubMed] [Google Scholar]
  5. Broze G. J., Jr, Majerus P. W. Purification and properties of human coagulation factor VII. J Biol Chem. 1980 Feb 25;255(4):1242–1247. [PubMed] [Google Scholar]
  6. Capecchi M. R. Altering the genome by homologous recombination. Science. 1989 Jun 16;244(4910):1288–1292. doi: 10.1126/science.2660260. [DOI] [PubMed] [Google Scholar]
  7. Chaing S., Clarke B., Sridhara S., Chu K., Friedman P., VanDusen W., Roberts H. R., Blajchman M., Monroe D. M., High K. A. Severe factor VII deficiency caused by mutations abolishing the cleavage site for activation and altering binding to tissue factor. Blood. 1994 Jun 15;83(12):3524–3535. [PubMed] [Google Scholar]
  8. Chen W. T. Membrane proteases: roles in tissue remodeling and tumour invasion. Curr Opin Cell Biol. 1992 Oct;4(5):802–809. doi: 10.1016/0955-0674(92)90103-j. [DOI] [PubMed] [Google Scholar]
  9. Davie E. W. Biochemical and molecular aspects of the coagulation cascade. Thromb Haemost. 1995 Jul;74(1):1–6. [PubMed] [Google Scholar]
  10. Degen S. J., Schaefer L. A., Jamison C. S., Grant S. G., Fitzgibbon J. J., Pai J. A., Chapman V. M., Elliott R. W. Characterization of the cDNA coding for mouse prothrombin and localization of the gene on mouse chromosome 2. DNA Cell Biol. 1990 Sep;9(7):487–498. doi: 10.1089/dna.1990.9.487. [DOI] [PubMed] [Google Scholar]
  11. Eichinger S., Mannucci P. M., Tradati F., Arbini A. A., Rosenberg R. D., Bauer K. A. Determinants of plasma factor VIIa levels in humans. Blood. 1995 Oct 15;86(8):3021–3025. [PubMed] [Google Scholar]
  12. Fenton J. W., 2nd Thrombin. Ann N Y Acad Sci. 1986;485:5–15. doi: 10.1111/j.1749-6632.1986.tb34563.x. [DOI] [PubMed] [Google Scholar]
  13. Hoffmann W. E., Everds N., Pignatello M., Solter P. F. Automated and semiautomated analysis of rat alkaline phosphatase isoenzymes. Toxicol Pathol. 1994 Nov-Dec;22(6):633–638. doi: 10.1177/019262339402200607. [DOI] [PubMed] [Google Scholar]
  14. Kazama Y., Hamamoto T., Foster D. C., Kisiel W. Hepsin, a putative membrane-associated serine protease, activates human factor VII and initiates a pathway of blood coagulation on the cell surface leading to thrombin formation. J Biol Chem. 1995 Jan 6;270(1):66–72. doi: 10.1074/jbc.270.1.66. [DOI] [PubMed] [Google Scholar]
  15. Kisiel W., Fujikawa K., Davie E. W. Activation of bovine factor VII (proconvertin) by factor XIIa (activated Hageman factor). Biochemistry. 1977 Sep 20;16(19):4189–4194. doi: 10.1021/bi00638a009. [DOI] [PubMed] [Google Scholar]
  16. Kraut J. Serine proteases: structure and mechanism of catalysis. Annu Rev Biochem. 1977;46:331–358. doi: 10.1146/annurev.bi.46.070177.001555. [DOI] [PubMed] [Google Scholar]
  17. Kurachi K., Torres-Rosado A., Tsuji A. Hepsin. Methods Enzymol. 1994;244:100–114. doi: 10.1016/0076-6879(94)44009-3. [DOI] [PubMed] [Google Scholar]
  18. Leytus S. P., Loeb K. R., Hagen F. S., Kurachi K., Davie E. W. A novel trypsin-like serine protease (hepsin) with a putative transmembrane domain expressed by human liver and hepatoma cells. Biochemistry. 1988 Feb 9;27(3):1067–1074. doi: 10.1021/bi00403a032. [DOI] [PubMed] [Google Scholar]
  19. Manfioletti G., Brancolini C., Avanzi G., Schneider C. The protein encoded by a growth arrest-specific gene (gas6) is a new member of the vitamin K-dependent proteins related to protein S, a negative coregulator in the blood coagulation cascade. Mol Cell Biol. 1993 Aug;13(8):4976–4985. doi: 10.1128/mcb.13.8.4976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mignatti P., Rifkin D. B. Biology and biochemistry of proteinases in tumor invasion. Physiol Rev. 1993 Jan;73(1):161–195. doi: 10.1152/physrev.1993.73.1.161. [DOI] [PubMed] [Google Scholar]
  21. Miller G. J., Martin J. C., Mitropoulos K. A., Esnouf M. P., Cooper J. A., Morrissey J. H., Howarth D. J., Tuddenham E. G. Activation of factor VII during alimentary lipemia occurs in healthy adults and patients with congenital factor XII or factor XI deficiency, but not in patients with factor IX deficiency. Blood. 1996 May 15;87(10):4187–4196. [PubMed] [Google Scholar]
  22. Morrissey J. H., Macik B. G., Neuenschwander P. F., Comp P. C. Quantitation of activated factor VII levels in plasma using a tissue factor mutant selectively deficient in promoting factor VII activation. Blood. 1993 Feb 1;81(3):734–744. [PubMed] [Google Scholar]
  23. Mortensen R. M., Conner D. A., Chao S., Geisterfer-Lowrance A. A., Seidman J. G. Production of homozygous mutant ES cells with a single targeting construct. Mol Cell Biol. 1992 May;12(5):2391–2395. doi: 10.1128/mcb.12.5.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nakamura T., Nishizawa T., Hagiya M., Seki T., Shimonishi M., Sugimura A., Tashiro K., Shimizu S. Molecular cloning and expression of human hepatocyte growth factor. Nature. 1989 Nov 23;342(6248):440–443. doi: 10.1038/342440a0. [DOI] [PubMed] [Google Scholar]
  25. Neuenschwander P. F., Morrissey J. H. Deletion of the membrane anchoring region of tissue factor abolishes autoactivation of factor VII but not cofactor function. Analysis of a mutant with a selective deficiency in activity. J Biol Chem. 1992 Jul 15;267(20):14477–14482. [PubMed] [Google Scholar]
  26. Neurath H. Evolution of proteolytic enzymes. Science. 1984 Apr 27;224(4647):350–357. doi: 10.1126/science.6369538. [DOI] [PubMed] [Google Scholar]
  27. Radcliffe R., Bagdasarian A., Colman R., Nemerson Y. Activation of bovine factor VII by hageman factor fragments. Blood. 1977 Oct;50(4):611–617. [PubMed] [Google Scholar]
  28. Schmidt C., Bladt F., Goedecke S., Brinkmann V., Zschiesche W., Sharpe M., Gherardi E., Birchmeier C. Scatter factor/hepatocyte growth factor is essential for liver development. Nature. 1995 Feb 23;373(6516):699–702. doi: 10.1038/373699a0. [DOI] [PubMed] [Google Scholar]
  29. Tanimoto H., Yan Y., Clarke J., Korourian S., Shigemasa K., Parmley T. H., Parham G. P., O'Brien T. J. Hepsin, a cell surface serine protease identified in hepatoma cells, is overexpressed in ovarian cancer. Cancer Res. 1997 Jul 15;57(14):2884–2887. [PubMed] [Google Scholar]
  30. Torres-Rosado A., O'Shea K. S., Tsuji A., Chou S. H., Kurachi K. Hepsin, a putative cell-surface serine protease, is required for mammalian cell growth. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7181–7185. doi: 10.1073/pnas.90.15.7181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tsuji A., Torres-Rosado A., Arai T., Chou S. H., Kurachi K. Characterization of hepsin, a membrane bound protease. Biomed Biochim Acta. 1991;50(4-6):791–793. [PubMed] [Google Scholar]
  32. Tsuji A., Torres-Rosado A., Arai T., Le Beau M. M., Lemons R. S., Chou S. H., Kurachi K. Hepsin, a cell membrane-associated protease. Characterization, tissue distribution, and gene localization. J Biol Chem. 1991 Sep 5;266(25):16948–16953. [PubMed] [Google Scholar]
  33. Uehara Y., Minowa O., Mori C., Shiota K., Kuno J., Noda T., Kitamura N. Placental defect and embryonic lethality in mice lacking hepatocyte growth factor/scatter factor. Nature. 1995 Feb 23;373(6516):702–705. doi: 10.1038/373702a0. [DOI] [PubMed] [Google Scholar]
  34. Wetsel R. A., Fleischer D. T., Haviland D. L. Deficiency of the murine fifth complement component (C5). A 2-base pair gene deletion in a 5'-exon. J Biol Chem. 1990 Feb 15;265(5):2435–2440. [PubMed] [Google Scholar]
  35. Wildgoose P., Berkner K. L., Kisiel W. Synthesis, purification, and characterization of an Arg152----Glu site-directed mutant of recombinant human blood clotting factor VII. Biochemistry. 1990 Apr 3;29(13):3413–3420. doi: 10.1021/bi00465a039. [DOI] [PubMed] [Google Scholar]
  36. Wildgoose P., Kisiel W. Activation of human factor VII by factors IXa and Xa on human bladder carcinoma cells. Blood. 1989 May 15;73(7):1888–1895. [PubMed] [Google Scholar]
  37. Wildgoose P., Nemerson Y., Hansen L. L., Nielsen F. E., Glazer S., Hedner U. Measurement of basal levels of factor VIIa in hemophilia A and B patients. Blood. 1992 Jul 1;80(1):25–28. [PubMed] [Google Scholar]
  38. Young G. P., Rose I. S., Cropper S., Seetharam S., Alpers D. H. Hepatic clearance of rat plasma intestinal alkaline phosphatase. Am J Physiol. 1984 Oct;247(4 Pt 1):G419–G426. doi: 10.1152/ajpgi.1984.247.4.G419. [DOI] [PubMed] [Google Scholar]
  39. Zhukov A., Hellman U., Ingelman-Sundberg M. Purification and characterization of hepsin from rat liver microsomes. Biochim Biophys Acta. 1997 Jan 4;1337(1):85–95. doi: 10.1016/s0167-4838(96)00152-5. [DOI] [PubMed] [Google Scholar]

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