Use of a bacteriophage-derived endo-N-acetylneuraminidase and an equine antipolysialyl antibody to characterize the polysialyl residues in salmonid fish egg polysialoglycoproteins. Substrate and immunospecificity studies.
Polysialoglycoproteins (PSGP), a class of glycoproteins containing oligo(poly)sialylglycan chains, are the major glycoprotein components in cortical alveoli of a number of Salmonidae fish eggs. Lake trout, Salvelinus namaycush, egg PSGP (PSGP(Sn)) differs from rainbow trout, Salmo gairdneri, egg PSGP (PSGP(Sg)) in its sialic acid composition; the former contains both N-acetyl- and N-glycolyl-D-neuraminic acid residues, designated Neu5Ac and Neu5Gc, while the latter contains only Neu5Gc residues. Fragmentation analysis of oligo(poly)sialyl chains in lake trout PSGP(Sn) has established that there are two distinct types of oligo(poly)sialyl structures in this PSGP molecule, namely alpha-2,8-linked oligo/poly(Neu5Ac) and alpha-2,8-linked oligo/poly(Neu5Gc). No hybrid structure having both Neu5Ac and Neu5Gc residues in the fragment oligosialic acids was detected. These two distinct PSGP preparations from eggs of lake trout and rainbow trout have been used to compare their immunoreactivity with anti-polysialyl antibodies (H.46) and sensitivity to a bacteriophage-derived (Escherichia coli K1F) endo-N-acetylneuraminidase (Endo-N). H.46 was found to cross-react only with lake trout PSGP(Sn) in immunodiffusion assays but not with rainbow trout PSGP(Sg), indicating that H.46 is a specific probe for alpha-2,8-linked poly(Neu5Ac) but not for poly(Neu5Gc). In contrast, Endo-N was found to catalyze the hydrolysis of both alpha-2,8-linked poly (Neu5Ac) and poly(Neu5Gc), so that this enzyme can be used as a diagnostic reagent for detecting both types of polysialic acids. H.46 was used in indirect immunofluorescence experiments to localize PSGP(Sn) in cortical alveoli isolated from lake trout eggs.
D000818:Animals; D000906:Antibodies; D003090:Coliphages / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D005455:Fluorescent Antibody Technique; D005779:Immunodiffusion; D009439:Neuraminidase; D010063:Ovum / Q000032:analysis*; D011134:Polysaccharides / Q000032:analysis*; D012487:Salmonidae; D012794:Sialic Acids / Q000032:analysis*; D012795:Sialoglycoproteins / Q000302:isolation & purification; D014337:Trout
D000906:Antibodies; D011134:Polysaccharides; D012794:Sialic Acids; D012795:Sialoglycoproteins; C021319:polysialic acid; D009439:Neuraminidase