Localization of the serine protease-binding sites in the collagen-like domain of mannose-binding protein: indirect effects of naturally occurring mutations on protease binding and activation.
Mutations in the collagen-like domain of serum mannose-binding protein (MBP) interfere with the ability of the protein to initiate complement fixation through the MBP-associated serine proteases (MASPs). The resulting deficiency in the innate immune response leads to susceptibility to infections. Studies have been undertaken to define the region of MBP that interacts with MASPs and to determine how the naturally occurring mutations affect this interaction. Truncated and modified MBPs and synthetic peptides that represent segments of the collagen-like domain of MBP have been used to demonstrate that MASPs bind on the C-terminal side of the hinge region formed by an interruption in the Gly-X-Y repeat pattern of the collagen-like domain. The binding sites for MASP-2 and for MASP-1 and -3 overlap but are not identical. The two most common naturally occurring mutations in MBP result in substitution of acidic amino acids for glycine residues in Gly-X-Y triplets on the N-terminal side of the hinge. Circular dichroism analysis and differential scanning calorimetry demonstrate that the triple helical structure of the collagen-like domain is largely intact in the mutant proteins, but it is more easily unfolded than in wild-type MBP. Thus, the effect of the mutations is to destabilize the collagen-like domain, indirectly disrupting the binding sites for MASPs. In addition, at least one of the mutations has a further effect on the ability of MBP to activate MASPs.
Wallis
Russell
R
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.
Shaw
Jonathan M
JM
Uitdehaag
Joost
J
Chen
Ce-Belle
CB
Torgersen
Dawn
D
Drickamer
Kurt
K
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D003094:Collagen / Q000737:chemistry*; D003165:Complement System Proteins / Q000378:metabolism; D037601:Mannose-Binding Lectin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D050606:Mannose-Binding Protein-Associated Serine Proteases; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017434:Protein Structure, Tertiary; D051381:Rats; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism*
D037601:Mannose-Binding Lectin; C445599:Mbl2 protein, rat; C069858:mannose binding protein A; D003094:Collagen; D003165:Complement System Proteins; D050606:Mannose-Binding Protein-Associated Serine Proteases; D012697:Serine Endopeptidases