Alpha-(2----8)-polysialic acid immunoreactivity in voltage-sensitive sodium channel of eel electric organ.
The voltage-sensitive sodium channel from eel electroplax is formed of a polypeptide of 208,321 Da, to which is attached ca. 85 kDa of carbohydrate. Sialic acid is a prominent constituent, contributing ca. 113 negative charges to the protein surface. We here demonstrate that antibodies raised against the bacterial antigen alpha-(2----8)-polysialic acid, specific for polymers of ten or more consecutive sialic acid residues, react specifically and with high affinity to the electroplax sodium channel. In extracts of electroplax membranes, the sodium channel is the only protein that demonstrates this immunoreactivity, suggesting the presence of a polysialosyl-sialyltransferase specifically committed to this unique post-translational modification of the sodium channel. Polysialic acid is rare in vertebrates, having previously been found only associated with neural-cell adhesion molecules, present in the developing neuromuscular system. The other prominent source is the capsular polysaccharide of highly pathogenic meningitis bacteria. Antibodies to the bacterial antigen thus provide highly specific affinity markers for the sodium channel. The high avidity of these antibodies and the ratio of sialic acid residues to consensus glycosylation sites suggest that the terminal chains are well over ten sialosyl residues in length, potentially extending 10-30 nm into the extracellular environment.
James
W M
WM
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06510.
Agnew
W S
WS
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004557:Electric Organ / Q000032:analysis*; D004593:Electrophorus; D007473:Ion Channels / Q000032:analysis*; D011134:Polysaccharides / Q000302:isolation & purification; D012794:Sialic Acids / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007473:Ion Channels; D011134:Polysaccharides; D012794:Sialic Acids; C021319:polysialic acid