title: Heterocomplex formation between MBL/ficolin/CL-11-associated serine protease-1 and -3 and MBL/ficolin/CL-11-associated protein-1.
abstract: The activity of the complement system is tightly controlled by many fluid-phase and tissue-bound regulators. Mannose-binding lectin (MBL)/ficolin/collectin-11-associated protein-1 (MAP-1) is a recently discovered plasma protein that acts as an upstream inhibitor of the lectin complement pathway (LCP). It has previously been shown that MAP-1 can compete with the MBL/ficolin/collectin-11-associated serine proteases (MASPs) in binding to MBL and the ficolins. However, this mechanism may only partly explain the inhibitory complement effect of MAP-1. We hypothesized that MAP-1 is also involved in heterocomplex formation with the MASPs thereby breaking the stoichiometry of the activation complexes of the LCP, which could represent an alternative mechanism of MAP-1-mediated complement inhibition. We assessed the heterocomplex formation with ELISA, size-exclusion chromatography, and immunoblotting using both recombinant proteins and serum/plasma. We found that rMAP-1 can engage in heterocomplexes with rMASP-1 and rMASP-3 in a calcium-dependent manner. Moreover, we discovered that rMASP-1 and rMASP-3 also form heterocomplexes under these conditions. Complexes containing both MAP-1 and MASP-1 or -3 were detected in normal human serum and plasma, and depletion of the LCP recognition molecules from ficolin-3-deficient human serum showed that free circulating heterocomplexes also exist in the blood, although the major part appears to be associated with the LCP recognition molecules. Altogether, these findings suggest that MASPs can associate in various combinations and bring new perspectives to the complexity of lectin pathway-driven complement activation.
authors.affiliation: Laboratory of Molecular Medicine, Department of Clinical Immunology, Faculty of Health and Medical Sciences, Rigshospitalet, University of Copenhagen, DK 2100 Copenhagen, Denmark.
authors.forename: Anne
authors.initials: A
authors.lastname: Rosbjerg
authors.forename: Lea
authors.initials: L
authors.lastname: Munthe-Fog
authors.forename: Peter
authors.initials: P
authors.lastname: Garred
authors.forename: Mikkel-Ole
authors.initials: MO
authors.lastname: Skjoedt
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D018920:Coculture Techniques; D003167:Complement Activation / Q000502:physiology*; D037582:Complement Pathway, Mannose-Binding Lectin / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D037102:Lectins / Q000378:metabolism; D037241:Mannose-Binding Lectins / Q000378:metabolism; D050606:Mannose-Binding Protein-Associated Serine Proteases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D000097604:Ficolins
chemical_list: D037102:Lectins; D037241:Mannose-Binding Lectins; D050606:Mannose-Binding Protein-Associated Serine Proteases; D011994:Recombinant Proteins; D000097604:Ficolins; C084067:MASP1 protein, human