Immunogenetics

, Volume 57, Issue 11, pp 837–844

Carbohydrate-binding specificities of mouse ficolin A, a splicing variant of ficolin A and ficolin B and their complex formation with MASP-2 and sMAP

  • Y. Endo
  • N. Nakazawa
  • Y. Liu
  • D. Iwaki
  • M. Takahashi
  • T. Fujita
  • M. Nakata
  • M. Matsushita
Original Paper

Abstract

Ficolins are a group of proteins mainly consisting of collagen-like and fibrinogen-like domains and are thought to play a role in innate immunity via their carbohydrate-binding activities. Two types of ficolins have been identified in mice, ficolin A, and ficolin B. However, their structure and function are not fully understood. In this study, we isolated the cDNA encoding a novel variant of ficolin A having a shorter collagen-like domain and a longer gap sequence, which was generated from the ficolin A gene by alternative splicing. We delineated the structure and function of mouse ficolins, including this splicing variant, by preparing the respective recombinants. Recombinant ficolin A, its splicing variant, and ficolin B showed multimeric structures and revealed binding to both N-acetylglucosamine and N-acetylgalactosamine. Interestingly, ficolin B specifically recognized sialic acid residues. Ficolin A and its variant, but not ficolin B, bound to mannose-binding lectin (MBL)-associated serine protease-2 (Masp-2) and small MBL-associated protein (smap), and the resulting complexes showed a potent complement activating capacity. In addition, smap competed with Masp-2 in association with ficolin A and its variant, and inhibited the complement activation by the ficolin A (or ficolin A variant)/MASP-2 complex, indicating its regulatory role in the lectin pathway. These results suggest that ficolin A and its variant function as recognition molecules of the lectin pathway, and ficolin B plays a distinct role through its unique carbohydrate-binding specificity.

Keywords

Ficolin Mannose-binding lectin (MBL) MBL-associated serine protease (MASP) Lectin pathway Host defense Alternative splicing 

References

  1. Aikawa M, Yae Y, Sugimoto R, Suzuki SO, Iwaki T, Izuhara K, Hamasaki N (1999) Hakata antigen, a new member of the ficolin/opsonin P35 family, is a novel human lectin secreted into bronchus/alveolus and bile. J Histochem Cytochem 47:777–785PubMedGoogle Scholar
  2. Aoyagi Y, Adderson EE, Min JG, Matsushita M, Fujita Takahashi TS, Okuwaki Y, Bohnsack JF (2005) Role of L-ficolin/mannose-binding lectin-associated serine protease complexes in the opsonophagocytosis of type III group B streptococci. J Immunol 174:418–425PubMedGoogle Scholar
  3. Dahl MR, Thiel S, Matsushita M, Fujita T, Willis AC, Christensen T, Vorup-Jensen T, Jensenius JC (2001) MASP-3 and its association with distinct complexes of the mannan-binding lectin complement activation pathway. Immunity 15:127–35CrossRefPubMedGoogle Scholar
  4. Edgar PF (1995) Hucolin, a new corticosteroid-binding protein from human plasma with structural similarity to ficolins, transforming growth factor-β1-binding protein. FEBS Lett 375:159–161CrossRefPubMedGoogle Scholar
  5. Endo Y, Sato Y, Matsushita M, Fujita T (1996) Cloning and characterization of the human lectin P35 gene and its related gene. Genomics 36:515–521CrossRefPubMedGoogle Scholar
  6. Endo Y, Takahashi M, Nakao M, Saiga H, Sekine H, Matsushita M, Nonaka M, Fujita T (1998) Two lineages of mannose-binding lectin-associated serine protease (MASP) in vertebrates. J Immunol 161:4924–4930PubMedGoogle Scholar
  7. Endo Y, Nonaka M, Saiga H, Kakinuma Y, Matsushita A, Takahashi M, Matsushita M, Fujita T (2003) Origin of mannose-binding lectin-associated serine protease (MASP)-1 and MASP-3 involved in the lectin complement pathway traced back to the invertebrate, amphioxus. J Immunol 170:4701–4707PubMedGoogle Scholar
  8. Endo Y, Liu Y, Kanno K, Takahashi M, Matsushita M, Fujita T (2004) Identification of the mouse H-ficolin gene as a pseudogene and orthology between mouse ficolins A/B and human L-/M-ficolins. Genomics 84:737–744CrossRefPubMedGoogle Scholar
  9. Fujimori Y, Harumiya S, Fukumoto Y, Miura Y, Yagasaki K, Tachikawa H, Fujimoto D (1998) Molecular cloning and characterization of mouse ficolin-A. Biochem Biophys Res Commun 244:796–800CrossRefPubMedGoogle Scholar
  10. Fujita T (2002) Evolution of the lectin-complement pathway and its role in innate immunity. Nat Rev Immunol 2:346–353CrossRefPubMedGoogle Scholar
  11. Harumiya S, Omori A, Sugimura T, Fukumoto Y, Tachikawa H, Fujimoto D (1995) EBP-37, a new elastin-binding protein in human placenta: structural similarity to ficolins, transforming growth factor-β1-binding proteins. J Biochem 117:1029–1035PubMedGoogle Scholar
  12. Ichijo H, Rönnstrand L, Miyagawa K, Ohashi H, Helden CH, Miyazono K (1991) Purification of transforming growth factor-β1 binding proteins from porcine uterus membranes. J Biol Chem 266:22459–22464PubMedGoogle Scholar
  13. Ichijo H, Hellmanm U, Wernstedtm C, Gonez LJ, Claesson-Welsh L, Helden CH, Miyazono K (1993) Molecular cloning and characterization of ficolin, a multimeric protein with fibrinogen- and collagen-like domains. J Biol Chem 268:14505–14513PubMedGoogle Scholar
  14. Iwaki D, Fujita T (2005) Production and purification of recombinants of mouse MASP-2 and sMAP. J Endotoxin Res 11:47–50CrossRefPubMedGoogle Scholar
  15. Kakinuma Y, Endo Y, Takahashi M, Nakata M, Matsushita M, Takenoshita S, Fujita T (2003) Molecular cloning and characterization of novel ficolins from Xenopus laevis. Immunogenetics 55:29–37PubMedGoogle Scholar
  16. Kenjo A, Takahashi M, Matsushita M, Endo Y, Nakata M, Mizuochi T, Fujita T (2001) Cloning and characterization of novel ficolins from the solitary ascidian, Halocynthia roretzi. J Biol Chem 276:19959–19965CrossRefPubMedGoogle Scholar
  17. Krarup A, Thiel S, Hansen A, Fujita T, Jensenius JC (2004) L-ficolin is a pattern recognition molecule specific for acetyl group. J Biol Chem 279:47513–47519CrossRefPubMedGoogle Scholar
  18. Kuge S, Ihara S, Watanabe E, Watanabe M, Takishima K, Suga T, Mamiya G, Kawakami M (1992) cDNAs and deduced amino acid sequences of subunits in the binding component of mouse bactericidal factor, Ra-reactive factor: similarity to mannose-binding proteins. Biochemistry 31:6943–6950CrossRefPubMedGoogle Scholar
  19. Le Y, Lee SH, Kon OL, Lu J (1998) Human L-ficolin: plasma levels, sugar specificity, and assignment of its lectin activity to the fibrinogen-like (FBG) domain. FEBS Lett 425:367–370CrossRefPubMedGoogle Scholar
  20. Liu Y, Endo Y, Homma S, Kanno K, Yaginuma H, Fujita T (2005a) Ficolin A and ficolin B are expressed in distinct ontogenic patterns and cell types in the mouse. Mol Immunol 42:1265–1273CrossRefPubMedGoogle Scholar
  21. Liu Y, Endo Y, Iwaki D, Nakata M, Matsushita M, Wada I, Inoue K, Munakata M, Fujita T (2005b) Human M-ficolin is a secretory protein that activates the lectin complement pathway. J Immunol 175:3150–3156PubMedGoogle Scholar
  22. Lu J, Tay PN, Kon OL, Reid KBM (1996) Human ficolin: cDNA cloning, demonstration of peripheral blood leucocytes as the major site of synthesis and assignment of the gene to chromosome 9. Biochem J 313:473–478PubMedGoogle Scholar
  23. Matsushita M, Ezekowitz RA, Fujita T (1995) The Gly-54→Asp allelic form of human mannose-binding protein (MBP) fails to bind MBP-associated serine protease. Biochem J 311:1021–1023PubMedGoogle Scholar
  24. Matsushita M, Endo Y, Taira S, Sato Y, Fujita T, Ichikawa N, Nakata M, Mizuochi T (1996) A novel human serum lectin with collagen- and fibrinogen-like domains that functions as an opsonin. J Biol Chem 271:2448–2454CrossRefPubMedGoogle Scholar
  25. Matsushita M, Endo Y, Fujita T (2000) Complement activation complex of ficolin and mannose-binding lectin-associated serine protease. J Immunol 164:2281–2284PubMedGoogle Scholar
  26. Matsushita M, Kuraya M, Hamasaki N, Tsujimura M, Shirai H, Fujita T (2002) Activation of the lectin complement pathway by H-ficolin (Hakata antigen). J Immunol 168:3502–3506PubMedGoogle Scholar
  27. Ohashi T, Erickson HP (1998) Oligomeric structure and tissue distribution of ficolins from mouse, pig and human. Arch Biochem Biophys 360:223–232CrossRefPubMedGoogle Scholar
  28. Sastry K, Zahedi K, Lelias JM, Whitehead AS, Ezekowitz RA (1991) Molecular characterization of the mouse mannose-binding proteins. The mannose-binding protein A but not C is an acute phase reactant. J Immunol 147:692–697PubMedGoogle Scholar
  29. Sotiropoulou G, Kono M, Anisowicz A, Stenman G, Tsuji S, Sager R (2002) Identification and functional characterization of a human GalNAc[alpha]2,6-sialyltransferase with altered expression in breast cancer. Mol Med 8:42–55PubMedGoogle Scholar
  30. Stover CM, Thiel S, Thelen M, Lynch NJ, Vorup-Jensen T, Jensenius JC, Schwaeble WJ (1999) Two constituents of the initiation complex of the mannan-binding lectin activation pathway of complement are encoded by a single structural gene. J Immunol 162:3481–3490PubMedGoogle Scholar
  31. Sugimoto R, Yae Y, Aikawa M, Kitajima S, Shibata Y, Sato H, Hirata J, Okochi K, Izuhara K, Hamasaki N (1998) Cloning and characterization of the Hakata antigen, a member of the ficolin/opsonin p35 lectin family. J Biol Chem 273:20721–20727CrossRefPubMedGoogle Scholar
  32. Takahashi M, Endo Y, Fujita T, Matsushita M (1999) A truncated form of mannose-binding lectin-associated serine protease (MASP)-2 expressed by alternative polyadenylation is a component of the lectin complement pathway. Int Immunol 11:859–863CrossRefPubMedGoogle Scholar
  33. Takayama Y, Takada F, Takahashi A, Kawakami M (1994) A 100-kDa protein in the C4-activating component of Ra-reactive factor is a new serine protease having module organization similar to C1r and C1s. J Immunol 152:2308–2316PubMedGoogle Scholar
  34. Tsujimura M, Miyazaki T, Kojima E, Sagara Y, Shiraki H, Okouchi K, Maeda Y (2002) Serum concentration of Hakata antigen, a member of the ficolin, is linked with inhibition of Aerococcus virdans growth. Clin Chim Acta 325:139–146CrossRefPubMedGoogle Scholar
  35. Wallis R, Shaw JM, Uitdehaag J, Chen C-B, Torgersen D, Drickmer K (2004) Localization of the serine protease-binding sites in the collage-like domain of mannose-binding protein. J Biol Chem 279:14065–14073CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Y. Endo
    • 1
  • N. Nakazawa
    • 1
  • Y. Liu
    • 1
  • D. Iwaki
    • 1
  • M. Takahashi
    • 1
  • T. Fujita
    • 1
  • M. Nakata
    • 2
  • M. Matsushita
    • 2
  1. 1.Departmant of ImmunologyFukushima Medical University School of MedicineFukushimaJapan
  2. 2.Department of Applied Biochemistry and Institute of GlycotechnologyTokai UniversityHiratsukaJapan

Personalised recommendations