The major Kurloff cell glycoproteins: lectin affinities, glycosidase susceptibilities and relationship with the sialylated acid phosphatases of the Kurloff body.
Urea-soluble fractions from purified Kurloff cells (KC) were analysed by affinoblotting. Lectin reactivities were quasi-exclusively confined to the 30-35 kDa major glycoproteins (mGPs) (responsible for the PAS positivity of the Kurloff body) with strong affinities for Canavalia ensiformis lectin, Phaseolus vulgaris erythroagglutinin and Sambucus nigra (SNA), Pisum sativum, Triticum vulgaris and Ulex europeus agglutinins. These data were consistent with the presence, among the KC mGPs, of large amounts of complex or hybrid N-glycosylproteins, in particular with Neu5Ac alpha 2,6Gal/GalNAc sequences, fucosyl residues and bisected residues. Their oligosaccharide sequences belong to more than one class, since some of these lectin reactivities had to be borne by distinct N-linked oligosaccharide chains. Before further analysis, KC mGPs were separated from other highly anionic glycoconjugates, by DEAE-cellulose chromatography. Their abundant potential RCA-binding sites masked by sialic acid were then revealed after neuraminidase (sialidase) or dilute acid pre-treatment. In remaining consistent with their lectin affinities, some KC mGPs were found to be PNGase F sensitive, while, either desialylated or not, they were all O-glycanase insensitive. Finally, by combined zymography and affinoblotting, the SNA-reactive fraction of KC mGPs was shown to correspond to denatured forms of the two zymographic size populations (190 kDa and 500 kDa) of KC acid phosphatases.
Landemore
G
G
Laboratoire d'Histologie, UFR de Médecine, Université de Caen, CHU Côte de Nacre, France.
Oulhaj
N
N
Letaïef
S E
SE
Izard
J
J
D000135:Acid Phosphatase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002240:Carbohydrate Sequence; D002469:Cell Separation / Q000379:methods; D002477:Cells / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006026:Glycoside Hydrolases; D006168:Guinea Pigs; D037102:Lectins; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012794:Sialic Acids / Q000032:analysis*; D013154:Spleen / Q000166:cytology* / Q000378:metabolism
D006023:Glycoproteins; D037102:Lectins; D012794:Sialic Acids; D000135:Acid Phosphatase; D006026:Glycoside Hydrolases