Binding of T-antigen-bearing neoglycoprotein and peanut agglutinin to cultured tumor cells and breast carcinomas.
Chemical conjugation of appropriate carbohydrate ligands to an inert labeled carrier renders probes available to screen for the presence of respective binding sites. A set with a certain plant lectin and a suitable neoglycoprotein can thus determine complementary parts of a potentially relevant glycobiological interaction system. Owing to the interest in the peanut agglutinin-reactive T-antigen, we performed chemical synthesis of the respective disaccharide structure to serve as glycohistochemical ligand and established refinements of the synthetic patway. Coupling of the derivatized monomers had to be performed in the presence of sodium sulfate for optimal results. Complete removal of the protective groups from the p-nitrophenyl derivative of the N-acetylgalactosamine moiety was achieved under mild conditions with 2,3-dichloro-5,6-dicyanobenzoquinone without affecting any other functional groups. Specific binding sites for the synthetic neoglycoprotein as well as for the plant lectin were demonstrated in cell lines of human breast carcinoma colon adenocarcinoma, and erythroleukemia. ABC reagents in conjunction with DAB as peroxidase substrate were used to visualize specific binding sites. Binding complied with the accepted criteria for specificity. Moreover, carbohydrate-specific binding sites were detected in sections of nine out of 14 cases with malignant breast lesions. The percentage of positive tumor cells with both neoglycoprotein and lectin was similar in each of the individual sections, regardless of quantitative variations between cases, lectin staining intensity often being more pronounced. The reactivity pattern in sections of primary and metastatic lesions was not significantly correlated with the lymph node status. This study emphasized that custom synthesis of saccharides and histochemical application of the resulting neoglycoprotein has a remarkable potential for complementary assessment of endogenous binding sites for carbohydrate structures, localized by external tools such as plant lectins, as a step to elucidate the importance of a putative proteincarbohydrate interaction.
Gabius
H J
HJ
Max-Planck-Institut für Experimentelle Medizin, Abteilung Chemie Göttingen, Federal Republic of Germany.
Schröter
C
C
Gabius
S
S
Brinck
U
U
Tietze
L F
LF
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology*; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism*; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism / Q000473:pathology*; D019887:Peanut Agglutinin; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure
D000945:Antigens, Differentiation, T-Lymphocyte; D015295:Antigens, Tumor-Associated, Carbohydrate; D006023:Glycoproteins; D037102:Lectins; D019887:Peanut Agglutinin; D011948:Receptors, Antigen, T-Cell