Characterization of the interaction between L-ficolin/P35 and mannan-binding lectin-associated serine proteases-1 and -2

S. Cseh, Loanys Vera, Misao Matsushita, Teizo Fujita, Gérard J. Arlaud, Nicole M. Thielens

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Abstract

Ficolins are oligomeric lectins comprising a collagen-like and a fibrinogen-like domain, with a binding specificity for N-acetylglucosamine. It has been reported recently that L-ficolin/P35 associates with mannan-binding lectin (MBL)-associated serine proteases (MASP-1 and -2) and MBL-associated protein 19 (MApl9) in serum and forms complexes able to activate complement. Using surface plasmon resonance spectroscopy we have shown that recombinant MASP-1 and -2, their N-terminal CUB1 (module originally found in complement proteins Clr/Cls, Uegf, and bone morphogenetic protein-1) -epidermal growth factor (EGF)-CUB2 and CUB1-EGF segments, and MApl9 bind to immobilized L-ficolin/P35 in the presence of Ca2+ ions. Comparable Kd values were obtained for the full-length proteases and their CUB1-EGF-CUB2 segments (9.2 and 10 nM for MASP-1 and 4.6 and 5.4 nM for MASP-2, respectively), whereas higher values were obtained for the CUB1-EGF segments (26.7, 15.6, and 14.3 nM for MASP-1, MASP-2, and MApl9). These values are in the same range as those determined for the interaction of these proteins with MBL. Binding was Ca2+ dependent and was only partly sensitive to EDTA for MASP-1, MASP-2, and MASP-2 CUB1-EGF-CUB2. Half-maximal binding was obtained at comparable Ca2+ concentrations for MASP-1 and MASP-2 (0.45 and 0.47 μM, respectively), their CUB1-EGF-CUB2 segments (0.37 and 0.72 μM), and their CUB1-EGF segments (0.31 and 0.79 μM). These values are lower than those determined in the case of MBL, indicating a difference between MBL and L-ficolin/P35 with respect to the Ca2+ dependence of their interaction with the MASPs. Preincubation of the MASPs with soluble MBL inhibited subsequent binding to immobilized L-ficolin/P35 and, conversely, suggesting that these lectins compete with each other for binding to the MASPs in vivo.

Original languageEnglish
Pages (from-to)5735-5743
Number of pages9
JournalJournal of Immunology
Volume169
Issue number10
Publication statusPublished - Nov 15 2002

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Mannose-Binding Protein-Associated Serine Proteases
Epidermal Growth Factor
Mannose-Binding Lectin
ficolin
Lectins
Bone Morphogenetic Protein 1

ASJC Scopus subject areas

  • Immunology

Cite this

Cseh, S., Vera, L., Matsushita, M., Fujita, T., Arlaud, G. J., & Thielens, N. M. (2002). Characterization of the interaction between L-ficolin/P35 and mannan-binding lectin-associated serine proteases-1 and -2. Journal of Immunology, 169(10), 5735-5743.

Characterization of the interaction between L-ficolin/P35 and mannan-binding lectin-associated serine proteases-1 and -2. / Cseh, S.; Vera, Loanys; Matsushita, Misao; Fujita, Teizo; Arlaud, Gérard J.; Thielens, Nicole M.

In: Journal of Immunology, Vol. 169, No. 10, 15.11.2002, p. 5735-5743.

Research output: Contribution to journalArticle

Cseh, S, Vera, L, Matsushita, M, Fujita, T, Arlaud, GJ & Thielens, NM 2002, 'Characterization of the interaction between L-ficolin/P35 and mannan-binding lectin-associated serine proteases-1 and -2', Journal of Immunology, vol. 169, no. 10, pp. 5735-5743.
Cseh, S. ; Vera, Loanys ; Matsushita, Misao ; Fujita, Teizo ; Arlaud, Gérard J. ; Thielens, Nicole M. / Characterization of the interaction between L-ficolin/P35 and mannan-binding lectin-associated serine proteases-1 and -2. In: Journal of Immunology. 2002 ; Vol. 169, No. 10. pp. 5735-5743.
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abstract = "Ficolins are oligomeric lectins comprising a collagen-like and a fibrinogen-like domain, with a binding specificity for N-acetylglucosamine. It has been reported recently that L-ficolin/P35 associates with mannan-binding lectin (MBL)-associated serine proteases (MASP-1 and -2) and MBL-associated protein 19 (MApl9) in serum and forms complexes able to activate complement. Using surface plasmon resonance spectroscopy we have shown that recombinant MASP-1 and -2, their N-terminal CUB1 (module originally found in complement proteins Clr/Cls, Uegf, and bone morphogenetic protein-1) -epidermal growth factor (EGF)-CUB2 and CUB1-EGF segments, and MApl9 bind to immobilized L-ficolin/P35 in the presence of Ca2+ ions. Comparable Kd values were obtained for the full-length proteases and their CUB1-EGF-CUB2 segments (9.2 and 10 nM for MASP-1 and 4.6 and 5.4 nM for MASP-2, respectively), whereas higher values were obtained for the CUB1-EGF segments (26.7, 15.6, and 14.3 nM for MASP-1, MASP-2, and MApl9). These values are in the same range as those determined for the interaction of these proteins with MBL. Binding was Ca2+ dependent and was only partly sensitive to EDTA for MASP-1, MASP-2, and MASP-2 CUB1-EGF-CUB2. Half-maximal binding was obtained at comparable Ca2+ concentrations for MASP-1 and MASP-2 (0.45 and 0.47 μM, respectively), their CUB1-EGF-CUB2 segments (0.37 and 0.72 μM), and their CUB1-EGF segments (0.31 and 0.79 μM). These values are lower than those determined in the case of MBL, indicating a difference between MBL and L-ficolin/P35 with respect to the Ca2+ dependence of their interaction with the MASPs. Preincubation of the MASPs with soluble MBL inhibited subsequent binding to immobilized L-ficolin/P35 and, conversely, suggesting that these lectins compete with each other for binding to the MASPs in vivo.",
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