Identification of a Functionally Relevant Signal Peptide of Mouse Ficolin A

  • Kwon, Sang-Hoon (Department of Microbiology, College of Medicine, Hallym University) ;
  • Kim, Min-Soo (Department of Microbiology, College of Medicine, Hallym University) ;
  • Kim, Dong-Bum (Center for Medical Science Research, College of Medicine, Hallym University) ;
  • Lee, Keun-Wook (Center for Medical Science Research, College of Medicine, Hallym University) ;
  • Choi, Soo-Young (Department of Biomedical Science, Hallym University) ;
  • Park, Jin-Seu (Department of Biomedical Science, Hallym University) ;
  • Kim, Yeon-Hyang (Department of Biomedical Science, Hallym University) ;
  • Lee, Young-Hee (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Kwon, Hyung-Joo (Department of Microbiology, College of Medicine, Hallym University)
  • Published : 2007.07.31


Mouse ficolin A is a plasma protein with lectin activity, and plays a role in host defense by binding carbohydrates, especially GlcNAc, on microorganisms. The ficolin A subunit consists of an N-terminal signal peptide, a collagen-like domain, and a C-terminal fibrinogen-like domain. In this study, we show that ficolin A can be synthesized and oligomerized in a cell and secreted into culture medium. We also identify a functionally relevant signal peptide of ficolin A by using MS/MS analysis to determine the N-terminal sequence of secreted ficolin A. When the signal peptide of mouse ficolin A was fused with enhanced green fluorescent protein (EGFP), EGFP was released into HEK 293 cell medium, suggesting that the signal peptide can efficiently direct ficolin A secretion. Moreover, our results suggest that the signal peptide of ficolin A has potential application for the production of useful secretory proteins.


EGFP;Ficolin A;Fusion protein;Secretion;Signal peptide


  1. Aderem, A. and Ulevitch, R. J. (2000) Toll-like receptors in the induction of the innate immune response. Nature 406, 782-787.
  2. Dahl, M. R., Thiel, S., Matsushita, M., Fujita, T., Willis, A. C., Christensen, T., Vorup-Jensen, T. and Jensenius, J. C. (2001) MASP-3 and its association with distinct complexes of the mannan-binding lectin complement activation pathway. Immunity 15, 127-135.
  3. Endo, Y., Nakazawa, N., Liu, Y., Iwaki, D., Takahashi, M., Fujita, T., Nakata, M. and Matsushita, M. (2005) Carbohydratebinding specificities of mouse ficolin A, a splicing variant of ficolin A and ficolin B and their complex formation with MASP-2 and sMAP. Immunogenetics 57, 837-844.
  4. Endo, Y., Sato, Y., Matsushita, M. and Fujita, T. (1996) Cloning and characterization of the human lectin P35 gene and its related gene. Genomics 36, 515-521.
  5. Gierasch, L. M. (1989) Signal sequences. Biochemistry 28, 923-930.
  6. Fujimori Y., Harumiya, S., Fukumoto, Y., Miura, Y., Yagasaki, K., Tachikawa, H. and Fujimoto, D. (1998) Molecular cloning and characterization of mouse ficolin-A. Biochem. Biophys. Res. Commun. 244, 796-800.
  7. Fujita, T. Matsushita, M. and Endo, Y. (2004) The lectincomplement pathway - its role in innate immunity and evolution. Immunol. Rev. 198, 185-202.
  8. Holmskov, U., Thiel, S. and Jensenius, J. C. (2003) Collection and ficolins: humoral lectins of the innate immune defense. Annu. Rev. Immunol. 21, 547-578.
  9. Ichijo, H., Hellman, U., Wernstedt, C., Gonez, L. J., Claesson-Welsh, L., Heldin, C. H. and Miyazono, K. (1993) Molecular cloning and characterization of ficolin, a multimeric protein with fibrinogen- and collagen-like domains. J. Biol. Chem. 268, 14505-14513.
  10. Janeway, C. A. Jr. and Medzhitov, R. (1998) Introduction: the role of innate immunity in the adaptive immune response. Semin. Immunol. 10, 349-350.
  11. Kwon, H. J., Lee, K. W., Yu, S. H., Han, J. H. and Kim, D. S. (2003) $NF-{\kappa}B-dependent$ regulation of tumor necrosis $factor-{\alpha}$ gene expression by CpG-oligodeoxynucleotides. Biochem. Biophys. Res. Commun. 311, 129-138.
  12. Lee Y., Sohn, W. J., Kim, D. S. and Kwon, H. J. (2004) $NF-{\kappa}B$-and c-Jun-dependent regulation of human cytomegalovirus immediate-early gene enhuncer/promoter in response to lipopolysaccharie and bacterial CpG-oligodeoxynucleotides in macrophase cell in-RAW 264.7 Eur. J. Biochem. 271, 1094-1105.
  13. Matsushita, M., Endo, Y., Taira, S., Sato, Y., Fujita, T., Ichikawa, N., Nakata, M. and Mizuochi, T. (1996) A novel human serum lectin with collagen- and fibrinogen-like domains that functions as an opsonin. J. Biol. Chem. 271, 2448-2454.
  14. Matsushita, M. and Fujita, T. (1992) Activation of the classical complement pathway by mannose-binding protein in association with a novel C1s-like serine protease. J. Exp. Med. 176, 1497-1502.
  15. Matsushita, M. and Fujita, T. (2001) Ficolins and the lectin complement pathway. Immunol. Rev. 180, 78-85.
  16. Ohashi, T. and Erickson, H. P. (1998) Oligomeric structure and tissue distribution of ficolins from mouse, pig and human. Arch. Biochem. Biophys. 360, 223-232.
  17. Rothman, J. E. and Wieland, F. T. (1996) Protein sorting by transport vesicles. Science 272, 227-234.
  18. Roosi, V., Cseh, S., Bally, I., Thielens, N. M., Jensenius, J. C. and Arlaud, G. J. (2002) Substrate specificities of recombinant mannan-binding lectin-associated serine proteases-1 and -2. J. Biol. Chem. 276, 40880-40887.
  19. Roos, A., Bouwman, L. H., Munoz, J., Zuiverloon, T., Faber-Krol, M. C., Fallaux-van den Houten, F. C., Klar-Mohamad, N., Hack, C. E., Tilanus, M. G. and Daha, M. R. (2003) Functional characterization of the lectin pathway of complement in human serum. Mol. Immunol. 39, 655-668.
  20. Thiel, S., Vorup-Jensen, T., Stover, C. M., Schwaeble, W., Laursen, S. B., Poulsen, K., Willis, A. C., Eggleton, P., Hansen, S., Holmskov, U., Reid, K. B. and Jensenius, J. C. (1997) A second serine protease associated with mannanbinding lectin that activates complement. Nature 386, 506-510.
  21. Vorup-Jensen, T., Petersen, S. V., Hansen, A. G., Poulsen, K., Schwaeble, W., Sim, R. B., Reid, K. B., Davis, S. J., Thiel, S. and Jensenius, J. C. (2000) Distinct pathways of mannanbinding lectin (MBL)- and C1-complex autoactivation revealed by reconstitution of MBL with recombinant MBL-associated serine protease-2. J. Immunol. 165, 2093-2100.
  22. Schwaeble, W., Dahl, M. R., Thiel, S., Stover, C. M. and Jensenius, J. C. (2002) The mannan-binding lectin-associated serine proteases (MASPs) and Map19: four components of the lectin pathway activation complex encoded by two genes. Immunobiology 205, 455-466.
  23. Sugimoto, R., Yae, Y., Aikawa, M., Kitajima, S., Shibata, Y., Sato, H., Hirata, J., Okochi, K., Izuhara, K. and Hamasaki, N. (1998) Cloning and characterization of the Hakata antigen, a member of the ficolin/opsonin p35 lectin family. J. Biol. Chem. 273, 20721-20727.

Cited by

  1. L-Ficolin Binds to the Glycoproteins Hemagglutinin and Neuraminidase and Inhibits Influenza A Virus Infection Both in vitro and in vivo vol.4, pp.3, 2012,