PURIFICATION AND CHARACTERIZATION OF MANNAN-BINDING PROTEIN FROM MOUSE SERUM

被引:25
作者
HOLT, P
HOLMSKOV, U
THIEL, S
TEISNER, B
HOJRUP, P
JENSENIUS, JC
机构
[1] ODENSE UNIV,INST MED BIOL,DEPT MED MICROBIOL,ODENSE,DENMARK
[2] ODENSE UNIV,INST MOLEC BIOL,ODENSE,DENMARK
[3] UNIV AARHUS,INST MED MICROBIOL,DEPT IMMUNOL,AARHUS,DENMARK
[4] STATE SERUM INST,DIV IMMUNOL,COPENHAGEN,DENMARK
关键词
D O I
10.1111/j.1365-3083.1994.tb03361.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Mouse mannan-binding protein (MBP) was identified in serum by its Ca2+-dependent binding to mannan. On gel permeation chromatography, the protein eluted corresponding to a molecular weight of approximately 750 kDa. Analysed on SDS-PAGE under reducing conditions, the polypeptide showed an apparent molecular weight of 28 kDa, while several high molecular weight bands were seen under nonreducing conditions. The presence of collagen-like domains within the molecule was indicated by a high glycine content (14.9%) and substantiated by sensitivity to collagenase. Rabbit anti-mouse MBP antisera were raised. The concentration of MBP in serum from normal mice was measured by rocket immunoelectrophoresis and found to be from below 1 mu g/ml to 100 mu g/ml (average 50 mu g/ml, n = 60). The binding of mouse MBP to mannan could be inhibited by mono- and disaccharides in the following order of potency: L-fucose > D-mannose > N-acetyl-D-glucosamine > maltose > D-mannoheptulose > D-glucose > N-acetyl-D-mannosamine much greater than lactose > D-galactose much greater than N-acetyl-D-galactosamine. Mouse MBP was shown to activate the classical complement cascade after binding to mannan. The sequence of 14 NH2-terminal amino acid residues of the molecule showed 93% identity to rat MBP-A and complete identity to the translated cDNA sequences for mouse MBP-A and mouse Ra-reactive factor component P28b (RaRF P28b) published previously. The amino acid composition of mouse MBP showed a high degree of homology to MBPs from other species and mouse RaRF P28b.
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页码:202 / 208
页数:7
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