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- Title
Isolation and Characterization of Porcine Mannan-Binding Proteins of Different Size and Ultrastructure.
- Authors
Storgaard, P.; Nielsen, E. Holm; Andersen, O.; Skriver, E.; Mortensen, H.; Højrup, P.; Leslie, G.; Holmskov, U.; Svehag, S.-E.
- Abstract
The authors report on the purification and characterization of mannan-binding proteins (MBP) isolated from porcine serum. The MBPs were purified by use of PEG precipitation, affinity chromatography on mannan-Sepharose, protein A- and anti-porcine IgM-Sepharose followed by gel filtration. The MBP proteins were collagenase sensitive and showed γ1-γ2-electrophoretic mobility. The MBP designated pMBP-28 had a molecular mass of 28 kDa when analysed on SDS-PAGE under reducing conditions and eluted corresponding to a molecular mass of approximately 700 kDa on gel filtration chromatography. Electron micrograpbs of pMBP-28 revealed an oligomeric protein similar to rodent MBP-A and human MBP but with a predominance of penta- and hexameric molecules. Another protein designated pMBP-27 was composed of peptides of 27kDa and had an Mr of 300-350kDa on gel filtration chromatography. Electron microscopy of pMBP-27 showed dimer and trimer molecules; the trimers without distinct stalk regions. The N-terminal 26 (pMBP-27) and 24 (MBP-28) amino acid residues showed 54% and 58% identity with human MBP. pMBP-28 showed a higher degree of sequence similarity to rat and mouse MBP-A (60% identity) than to mouse and rat MBP-C (41-45% identity). Both pMBPs exhibited Ca2+-dependent binding to D-mannose immobilized on agarose but no significant binding to N-acetyl-D-glucosamine-or fucose-agarose. The results further suggested the presence of a third pMBP which copurified with pMBP-27 but this protein was not sequenced.
- Subjects
CHROMATOGRAPHIC analysis; GEL permeation chromatography; PROTEINS; COLLOIDS; POROUS materials; BIOMOLECULES; IMMUNOELECTROPHORESIS
- Publication
Scandinavian Journal of Immunology, 1996, Vol 43, Issue 3, p289
- ISSN
0300-9475
- Publication type
Article
- DOI
10.1046/j.1365-3083.1996.d01-39.x