Oligomeric structures required for complement activation of serum mannan-binding proteins.
Serum mannan-binding protein (S-MBP) comprises a series of homooligomers, and activates complement when it binds to appropriate carbohydrate ligands. In this study, the structural requirements necessary for complement activation were examined for rat, rabbit, and human S-MBPs. On SDS-PAGE under non-reducing conditions, the S-MBPs gave three major bands: large, middle, and small oligomers. Since three subunits (23-25 kDa) form a triple helix (the base structural unit) at the collagen-like domain within the S-MBP molecule, it was estimated that human and rabbit S-MBPs comprise a mixture of pentamers, tetramers, and trimers of the respective structural units. In contrast, rat S-MBP is composed of tetramers, trimers, and dimers. The large and middle oligomers were almost equal in their ability to activate complement, whereas the small oligomers had very low or no activity. Upon digestion with bacterial collagenase, the large and small oligomers were degraded almost completely. In contrast, the middle oligomers remained largely intact, and the surviving middle oligomers retained complete ability to activate complement. The degraded product, trimers of the carbohydrate recognition domain (CRD), did not show any complement activating activity. These data indicate that not only the structural integrity of the S-MBP collagen-like domain and CRD, but also a unique conformational structure present in the middle oligomers are critically important for carbohydrate-mediated complement activation.
Yokota
Y
Y
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University.
Arai
T
T
Kawasaki
T
T
D000818:Animals; D002352:Carrier Proteins / Q000097:blood* / Q000737:chemistry* / Q000302:isolation & purification; D002850:Chromatography, Gel; D037222:Collectins; D003167:Complement Activation; D006461:Hemolysis; D006801:Humans; D046911:Macromolecular Substances; D008351:Mannans / Q000378:metabolism*; D008970:Molecular Weight; D017433:Protein Structure, Secondary; D011817:Rabbits; D051381:Rats; D012441:Saccharomyces cerevisiae
D002352:Carrier Proteins; D037222:Collectins; D046911:Macromolecular Substances; D008351:Mannans