Tetanus toxin receptors on nerve cells contain a trypsin-sensitive component.
Cerebral neurons in monolayer cultures, subjected to 25 micrograms/ml trypsin, lose after 10 min about 43.5% and 40.5% of the ability to bind 125I-labeled tetanotoxin as measured at 0-4 degrees C and 37 degrees C respectively. These losses are maximal by 30 min and can be prevented by 1.5 mg/ml soybean trypsin inhibitor. Chymotrypsin but not collagenase or hyaluronidase is also effective in reducing binding of toxin to cells. The trypsin-insensitive toxin-binding activity can be further eliminated by treatment with sialidase or by cell extraction with methanol. Fixation of cells with 3.5% paraformaldehyde or 2% glutaraldehyde also results in a marked decrease of 52.4% and 25% respectively in the toxin-cell association. Methanol or sialidase but not trypsin removes the remaining binding activity. About one-third of the lipid-linked and protein-linked sialic acid is removed after sialidase treatment whereas 1% and 9.4% respectively are removed after trypsin treatment. The data are consistent with the possibility that, in addition to a sialic acid component, binding of tetanotoxin to nerve cells is facilitated by a trypsin-removable and formaldehyde-inactivated component. There was no evidence for a polypeptide to substitute gangliosides as receptors for tetanotoxin. On the contrary, solubility in organic solvents and interaction of the extracted products with labeled toxin remain the major proof that gangliosides are the putative receptors for tetanotoxin.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000494:pharmacology; D008055:Lipids / Q000302:isolation & purification; D008565:Membrane Proteins; D009439:Neuraminidase; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D011950:Receptors, Cholinergic / Q000378:metabolism*; D012995:Solubility; D014357:Trypsin
D003432:Cross-Linking Reagents; D008055:Lipids; D008565:Membrane Proteins; D011950:Receptors, Cholinergic; C028728:tetanus toxin receptor; D009439:Neuraminidase; D014357:Trypsin