Interaction of C1q and mannan-binding lectin (MBL) with C1r, C1s, MBL-associated serine proteases 1 and 2, and the MBL-associated protein MAp19.
Mannan-binding lectin (MBL) and C1q activate the complement cascade via attached serine proteases. The proteases C1r and C1s were initially discovered in a complex with C1q, whereas the MBL-associated serine proteases 1 and 2 (MASP-1 and -2) were discovered in a complex with MBL. There is controversy as to whether MBL can utilize C1r and C1s or, inversely, whether C1q can utilize MASP-1 and 2. Serum deficient in C1r produced no complement activation in IgG-coated microwells, whereas activation was seen in mannan-coated microwells. In serum, C1r and C1s were found to be associated only with C1q, whereas MASP-1, MASP-2, and a third protein, MAp19 (19-kDa MBL-associated protein), were found to be associated only with MBL. The bulk of MASP-1 and MAp19 was found in association with each other and was not bound to MBL or MASP-2. The interactions of MASP-1, MASP-2, and MAp19 with MBL differ from those of C1r and C1s with C1q in that both high salt concentrations and calcium chelation (EDTA) are required to fully dissociate the MASPs or MAp19 from MBL. In the presence of calcium, most of the MASP-1, MASP-2, and MAp19 emerged on gel-permeation chromatography as large complexes that were not associated with MBL, whereas in the presence of EDTA most of these components formed smaller complexes. Over 95% of the total MASPs and MAp19 found in serum are not complexed with MBL.
Thiel
S
S
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark. st@microbiology.au.dk
Petersen
S V
SV
Vorup-Jensen
T
T
Matsushita
M
M
Fujita
T
T
Stover
C M
CM
Schwaeble
W J
WJ
Jensenius
J C
JC
D002118:Calcium / Q000737:chemistry; D002352:Carrier Proteins / Q000097:blood / Q000276:immunology / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D037222:Collectins; D003172:Complement C1 / Q000378:metabolism*; D015922:Complement C1q / Q000276:immunology / Q000378:metabolism; D015923:Complement C1r / Q000378:metabolism; D003173:Complement C1s / Q000378:metabolism; D015935:Complement C4b / Q000378:metabolism; D004492:Edetic Acid / Q000737:chemistry; D006801:Humans; D007106:Immune Sera / Q000737:chemistry; D007074:Immunoglobulin G / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D008351:Mannans / Q000378:metabolism; D050606:Mannose-Binding Protein-Associated Serine Proteases; D009994:Osmolar Concentration; D011485:Protein Binding / Q000276:immunology; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*
D002352:Carrier Proteins; D037222:Collectins; D003172:Complement C1; D007106:Immune Sera; D007074:Immunoglobulin G; D037102:Lectins; D008351:Mannans; D015922:Complement C1q; D015935:Complement C4b; D004492:Edetic Acid; C105296:MASP2 protein, human; D050606:Mannose-Binding Protein-Associated Serine Proteases; D012697:Serine Endopeptidases; D015923:Complement C1r; D003173:Complement C1s; D002118:Calcium