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Published on: February 11, 2009
Characterization of a 180 kDa molecule apparently reactive with recombinant L-selectin
H Kawashima1, N Watanabe, Y F Li
1Department of Bioregulation, Osaka University Medical School, Suita, Japan.
Glycoconjugate Journal
|April 1, 1997
Summary
A 180 kDa molecule (p180) in rat lymph nodes reacts with silkworm L-selectin but not COS-7 L-selectin. This indicates p180 recognizes silkworm-specific carbohydrates, not L-selectin itself, cautioning against using certain recombinant proteins.
Area of Science:
- Immunology
- Glycobiology
- Molecular Biology
Background:
- L-selectin plays a role in lymphocyte homing.
- Recombinant proteins are often used to study molecular interactions.
- Expression systems can influence protein glycosylation.
Purpose of the Study:
- To identify molecules in rat lymph nodes reactive with silkworm-derived L-selectin.
- To investigate the nature of the interaction between p180 and L-selectin.
- To assess the reliability of silkworm-expressed recombinant proteins for ligand detection.
Main Methods:
- Identification of a 180 kDa molecule (p180) in rat lymph nodes.
- Amino acid sequence analysis and homology search.
- Binding assays using silkworm and COS-7 derived L-selectin-IgG.
- Inhibition assays with alpha-methyl-D-mannoside.
- Immunohistochemical studies.
Main Results:
- p180 showed homology to the macrophage mannose receptor (MMR).
- p180 binding to silkworm L-selectin was Ca(2+)-dependent and inhibited by mannosides.
- p180 did not bind to COS-7 derived L-selectin.
- p180 was found in medullary macrophages, not high endothelial venules.
Conclusions:
- p180 interacts with silkworm-specific oligomannose oligosaccharides, not L-selectin's lectin activity.
- p180 is not a physiological ligand for L-selectin.
- Caution is advised when using silkworm-expressed recombinant proteins for detecting carbohydrate ligands.
