Glycopeptide-albumin derivative: it preparation and histochemical ligand properties

Carrier-immobilized mono- or disaccharides and other carbohydrate structures, derived by custom-made chemical synthesis, have already proven to be valuable ligands for localizing carbohydrate-binding proteins in tissue sections. Defined purified glycopeptides, as components of neoglycoproteins, offe...

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Bibliographic Details
Published in:The Histochemical journal 1991-07, Vol.23 (7), p.303-311
Main Authors: Gabius, H J, Brinck, U, Lüsebrink, T, Ciesiolka, T, Gabius, S
Format: Article
Language:English
Subjects:
Asparagine - analogs & derivatives
Asparagine - chemistry
B-Lymphocytes - chemistry
Binding Sites
Biotin - chemistry
Breast Neoplasms - chemistry
Cell Line
Concanavalin A - chemistry
Cross-Linking Reagents
Electrophoresis, Polyacrylamide Gel
Histocytochemistry
Humans
Leukemia - metabolism
Mannose - metabolism
Oligosaccharides - chemistry
Pronase - metabolism
Serum Albumin
Serum Albumin, Bovine - chemistry
Staining and Labeling
Succinimides - chemistry
T-Lymphocytes - chemistry
Tumor Cells, Cultured
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Summary:Carrier-immobilized mono- or disaccharides and other carbohydrate structures, derived by custom-made chemical synthesis, have already proven to be valuable ligands for localizing carbohydrate-binding proteins in tissue sections. Defined purified glycopeptides, as components of neoglycoproteins, offer the possibility of increasing their structural complexity and, thereby, their receptor selectivity. To test the feasibility of this approach, the glycopeptide man6-glcNAc2-asparagine derived from ovalbumin was purified after pronase digestion. It was coupled to bovine serum albumin as carrier protein with the homobifunctional linking agent bis-(sulphosuccinimidyl)suberate to yield the diglycosylated concanavalin A-reactive product. Following biotinylation, it was used to detect mannose-specific binding sites in fixed cells of seven human leukemia or lymphoma lines and in fixed, paraffin-embedded sections of human breast cancer. In comparison to chemically mannosylated bovine serum albumin with ten sites of glycosylation or to ovalbumin, this derivative produced a similar pattern of reaction with a quantitatively lower extent of staining in most cases. Remarkably, the presence of potential endogenous ligands for the detected receptor sites was ascertained using the plant lectin concanavalin A. Thus, the conjugation of a purified, deliberately selected glycopeptide to a suitable carrier produces a histochemical tool for detecting glycopeptide-specific binding sites.
ISSN:0018-2214