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 language | English |
|---|---|
| Pages (from-to) | 5735-5743 |
| Number of pages | 9 |
| Journal | Journal of Immunology |
| Volume | 169 |
| Issue number | 10 |
| Publication status | Published - Nov 15 2002 |
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ASJC Scopus subject areas
- Immunology
Cite this
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 journal › Article
}
TY - JOUR
T1 - Characterization of the interaction between L-ficolin/P35 and mannan-binding lectin-associated serine proteases-1 and -2
AU - Cseh, S.
AU - Vera, Loanys
AU - Matsushita, Misao
AU - Fujita, Teizo
AU - Arlaud, Gérard J.
AU - Thielens, Nicole M.
PY - 2002/11/15
Y1 - 2002/11/15
N2 - 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.
AB - 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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M3 - Article
VL - 169
SP - 5735
EP - 5743
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 10
ER -