Skip to main content
SpringerLink
Log in

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

  • Original Paper
  • Published:
Immunogenetics Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • 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 47:777–785

    PubMed  Google Scholar 

  • 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 174:418–425

    PubMed  CAS  Google Scholar 

  • 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 15:127–35

    Article  PubMed  CAS  Google Scholar 

  • 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 375:159–161

    Article  PubMed  CAS  Google Scholar 

  • Endo Y, Sato Y, Matsushita M, Fujita T (1996) Cloning and characterization of the human lectin P35 gene and its related gene. Genomics 36:515–521

    Article  PubMed  CAS  Google Scholar 

  • 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 161:4924–4930

    PubMed  CAS  Google Scholar 

  • 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 170:4701–4707

    PubMed  CAS  Google Scholar 

  • 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 84:737–744

    Article  PubMed  CAS  Google Scholar 

  • 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 244:796–800

    Article  PubMed  CAS  Google Scholar 

  • Fujita T (2002) Evolution of the lectin-complement pathway and its role in innate immunity. Nat Rev Immunol 2:346–353

    Article  PubMed  CAS  Google Scholar 

  • 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 117:1029–1035

    PubMed  CAS  Google Scholar 

  • 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 266:22459–22464

    PubMed  CAS  Google Scholar 

  • 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 268:14505–14513

    PubMed  CAS  Google Scholar 

  • Iwaki D, Fujita T (2005) Production and purification of recombinants of mouse MASP-2 and sMAP. J Endotoxin Res 11:47–50

    Article  PubMed  CAS  Google Scholar 

  • 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 55:29–37

    PubMed  CAS  Google Scholar 

  • 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 276:19959–19965

    Article  PubMed  CAS  Google Scholar 

  • 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 279:47513–47519

    Article  PubMed  CAS  Google Scholar 

  • 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 31:6943–6950

    Article  PubMed  CAS  Google Scholar 

  • 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 425:367–370

    Article  PubMed  CAS  Google Scholar 

  • 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 42:1265–1273

    Article  PubMed  CAS  Google Scholar 

  • 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 175:3150–3156

    PubMed  CAS  Google Scholar 

  • 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 313:473–478

    PubMed  CAS  Google Scholar 

  • 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 311:1021–1023

    PubMed  CAS  Google Scholar 

  • 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 271:2448–2454

    Article  PubMed  CAS  Google Scholar 

  • Matsushita M, Endo Y, Fujita T (2000) Complement activation complex of ficolin and mannose-binding lectin-associated serine protease. J Immunol 164:2281–2284

    PubMed  CAS  Google Scholar 

  • 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 168:3502–3506

    PubMed  CAS  Google Scholar 

  • Ohashi T, Erickson HP (1998) Oligomeric structure and tissue distribution of ficolins from mouse, pig and human. Arch Biochem Biophys 360:223–232

    Article  PubMed  CAS  Google Scholar 

  • 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 147:692–697

    PubMed  CAS  Google Scholar 

  • 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 8:42–55

    PubMed  CAS  Google Scholar 

  • 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 162:3481–3490

    PubMed  CAS  Google Scholar 

  • 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 273:20721–20727

    Article  PubMed  CAS  Google Scholar 

  • 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 11:859–863

    Article  PubMed  CAS  Google Scholar 

  • 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 152:2308–2316

    PubMed  CAS  Google Scholar 

  • 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 325:139–146

    Article  PubMed  CAS  Google Scholar 

  • 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 279:14065–14073

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by grants-in-aid from the Ministry of Education, Science, Sports and Technology of Japan and from CREST, The Japan Science and Technology Agency.

Author information

Authors and Affiliations

  1. Departmant of Immunology, Fukushima Medical University School of Medicine, 1-Hikarigaoka, Fukushima, 960-1295, Japan

    Y. Endo, N. Nakazawa, Y. Liu, D. Iwaki, M. Takahashi & T. Fujita

  2. Department of Applied Biochemistry and Institute of Glycotechnology, Tokai University, Hiratsuka, Kanagawa, 259-1292, Japan

    M. Nakata & M. Matsushita

Authors
  1. Y. Endo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Nakazawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. Iwaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T. Fujita
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M. Nakata
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M. Matsushita
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Y. Endo.

Additional information

The nucleotide sequences reported in this paper have been submitted to the DDBJ, EMBL, and GenBank nucleotide sequence databases and have been assigned accession numbers AB222271 for ficolin A variant cDNA.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Endo, Y., Nakazawa, N., Liu, Y. et al. 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. Immunogenetics 57, 837–844 (2005). https://doi.org/10.1007/s00251-005-0058-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00251-005-0058-1

Keywords

Search

Navigation