Structure of the catalytic region of human complement protease C.hivin.1s: Study by chemical crosslinking and three-dimensional homology modeling

Rossi, Véronique; Gaboriaud, Christine; Lacroix, Monique; Ulrich, J.; Fontecilla‐Camps, Juan C.; Gagnon, Jean; Arlaud, Gérard J.

doi:10.1021/bi00022a004

Cited by 44 publications

(47 citation statements)

References 30 publications

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“…In contrast, the CUB1-EGF-CUB2 fragment of MASP-1 was produced at a satisfactory yield (ϳ10 g/ml), comparable to that obtained previously with the CUB1-EGF fragment of MASP-1 (21). As expected from the occurrence of two N-glycosylation sites at Asn 30 and Asn 159 (27), the CUB1-EGF-CUB2 fragment of MASP-1 was produced in a glycosylated form. The data obtained by mass spectrometry analysis were consistent with the presence of two short high mannose oligosaccharides and provided no evidence for distinct species bearing either one or two carbohydrates, as observed previously in the case of the CUB1-EGF fragment (21).…”

Section: Discussionsupporting

confidence: 79%

“…N-terminal sequence analyses were performed after SDS-PAGE and electrotransfer, using an Applied Biosystems model 477A protein sequencer (Foster City, CA) as described previously (30). Mass spectrometry analyses were performed using the matrix-assisted laser desorption ionization technique on a Voyager Elite XL instrument (Perseptive Biosystems, Cambridge, MA), under conditions described previously (31).…”

Section: Chemical Characterization Of the Recombinant Proteinsmentioning

confidence: 99%

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Characterization of the Interaction Between L-Ficolin/P35 and Mannan-Binding Lectin-Associated Serine Proteases-1 and -2

Cseh

Vera

Matsushita

et al. 2002

The Journal of Immunology

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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 (MAp19) 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 C1r/C1s, Uegf, and bone morphogenetic protein-1)-epidermal growth factor (EGF)-CUB2 and CUB1-EGF segments, and MAp19 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 MAp19). 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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Section: Discussionsupporting

confidence: 79%

Section: Chemical Characterization Of the Recombinant Proteinsmentioning

confidence: 99%

Characterization of the Interaction Between L-Ficolin/P35 and Mannan-Binding Lectin-Associated Serine Proteases-1 and -2

Cseh

Vera

Matsushita

et al. 2002

The Journal of Immunology

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“…N-terminal sequence analysis of the purified C1q globular domain was performed after SDS-PAGE and electrotransfer using an Applied Biosystems model 477 A protein sequencer as described previously [21].…”

Section: N-terminal Sequence Analysismentioning

confidence: 99%

Trimeric reassembly of the globular domain of human C1q

Tacnet

Cheong

Goeltz

et al. 2008

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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C1q is a versatile recognition protein which binds to a variety of targets and consequently triggers the classical pathway of complement. C1q is a hetero-trimer composed of three chains (A, B and C) arranged in three domains, a short N-terminal region, followed by a collagenous repeat domain that gives rise to the formation of (ABC) triple helices, each ending in a C-terminal hetero-trimeric globular domain, called gC1q, which is responsible for the recognition properties of C1q. The mechanism of the trimeric assembly of C1q and in particular the role of each domain in the process is unknown. Here, we have investigated if the gC1q domain was able to assemble into functional trimers, in vitro, in the absence of the collagenous domain, a motif known to promote obligatory trimers in other proteins. Acid-mediated gC1q protomers reassembled into functional trimers, once neutralized, indicating that it is the gC1q domain which possesses the information for trimerization. However, reassembly occurred after neutralization, only if the gC1q protomers had preserved a residual tertiary structure at the end of the acidic treatment. Thus, the collagenous domain of C1q might initialize the folding of the gC1q domain so that subsequent assembly of the entire molecule can occur.

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“…N-terminal Sequence Determination-N-terminal sequence analyses were performed after SDS-PAGE and electrotransfer, using an Applied Biosystems model 477A protein sequencer as described previously (38).…”

Section: Construction Of Expression Plasmids Containing Full-length Mmentioning

confidence: 99%

Substrate Specificities of Recombinant Mannan-binding Lectin-associated Serine Proteases-1 and -2

Rossi¹,

Cseh²,

Bally³

et al. 2001

Journal of Biological Chemistry

167

135

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Mannan-binding lectin (MBL)-associated serine proteases-1 and 2 (MASP-1 and MASP-2) are homologous modular proteases that each interact with MBL, an oligomeric serum lectin involved in innate immunity. To precisely determine their substrate specificity, human MASP-1 and MASP-2, and fragments from their catalytic regions were expressed using a baculovirus/insect cells system. Recombinant MASP-2 displayed a rather wide, C1s-like esterolytic activity, and specifically cleaved complement proteins C2 and C4, with relative efficiencies 3-and 23-fold higher, respectively, than human C1s. MASP-2 also showed very weak C3 cleaving activity. Recombinant MASP-1 had a lower and more restricted esterolytic activity. It showed marginal activity toward C2 and C3, and no activity on C4. The enzymic activity of both MASP-1 and MASP-2 was specifically titrated by C1 inhibitor, and abolished at a 1:1 C1 inhibitor:protease ratio. Taken together with previous findings, these and other data strongly support the hypothesis that MASP-2 is the protease that, in association with MBL, triggers complement activation via the MBL pathway, through combined self-activation and proteolytic properties devoted to C1r and C1s in the C1 complex. In view of the very low activity of MASP-1 on C3 and C2, our data raise questions about the implication of this protease in complement activation.Mannan-binding lectin (MBL) 1 is an oligomeric C-type lectin that recognizes arrays of neutral carbohydrates such as mannose and N-acetylglucosamine on the surface of pathogenic microorganisms (2). This selectivity endows MBL with the ability to discriminate self from infectious non-self, and confers this "ante-antibody" a major role in innate immunity, as underlined by numerous clinical reports indicating that MBL deficiency is linked with increased susceptibility to infectious diseases (3-5). In addition to its role as an opsonin (3), MBL has devised the ability to associate to several modular proteases termed MASPs (MBL-associated serine proteases) (6 -9). A single MASP entity was initially identified, and characterized as a protease with the ability to cleave complement proteins C4, C2, and C3 (7, 10). Further studies by Thiel et al. (6) revealed that MASP was indeed a mixture of two related but distinct proteases, MASP-1 and MASP-2, and that only the latter had the ability to cleave C4. A third protein component MAp19, arising from alternative splicing of the MASP-2 gene (11, 12), and very recently a further protease MASP-3 (13) were also shown to be associated with MBL.MASP-1 and MASP-2 show a domain organization identical to that of C1r and C1s, the enzymatic components of the C1 complex of complement (14), with an N-terminal CUB module (15) followed by an epidermal growth factor-like module, a second CUB module, two contiguous CCP modules (16), and a C-terminal chymotrypsin-like serine protease domain (see Fig. 1). By analogy with human C1s, it may be anticipated that the proteolytic activity and specificity of the MASPs is defined by the two CCP modules ...

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Structure of the catalytic region of human complement protease C.hivin.1s: Study by chemical crosslinking and three-dimensional homology modeling

Cited by 44 publications

References 30 publications

Characterization of the Interaction Between L-Ficolin/P35 and Mannan-Binding Lectin-Associated Serine Proteases-1 and -2

Characterization of the Interaction Between L-Ficolin/P35 and Mannan-Binding Lectin-Associated Serine Proteases-1 and -2

Trimeric reassembly of the globular domain of human C1q

Substrate Specificities of Recombinant Mannan-binding Lectin-associated Serine Proteases-1 and -2

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