Neuronal binding of tetanus toxin compared to its ganglioside binding fragment (H(c)).
The non-toxin 50 kD C-terminus peptide of the heavy chain of tetanus H(c) contains the ganglioside binding domain of tetanus toxin (TTX). H(c) retains much of the capacity of tetanus toxin for binding internalization and transport by neurons. For this reason tetanus H(c) has been studied as a vector for delivery of therapeutic proteins to neurons. We directly compared H(c) and TTX in the capacity to bind and be internalized by neurons by ELISA. Primary cultures of dissociated fetal cortical neurons were incubated with equimolar amounts of TTX or H(c). Neuronal associated tetanus protein was 4-8 fold greater on a molar basis with tetanus toxin compared to H(c) (1 h incubation). This increase in neuronal tetanus protein was evident with incubation in concentrations from 0.1 microM to 2 microM. There were greater amounts of TTX delivered to the cultured cells at both 0 degrees C (representing membrane bound tetanus protein) and 37 degrees C (bound and internalized tetanus protein). Unlike H(c), TTX showed significant continued accumulation of protein with increasing incubation durations. Neuronal associated TTX increased 2-3 fold over incubation times ranging from 1 to 8 h. Tetanus toxin appears to be clearly superior to the ganglioside binding fragment (H(c)) in the capacity for neuronal binding and internalization. Atoxic tetanus proteins containing additional molecular domains as well as H(c) may be more suitable vectors for linkage with therapeutic proteins and delivery to neurons.
Fishman
P S
PS
Neurology Service, Baltimore VA Medical Center and the Department of Neurology, University of Maryland, School of Medicine, Baltimore, USA. pfishman@umaryland.edu
Parks
D A
DA
Patwardhan
A J
AJ
Matthews
C C
CC
D000818:Animals; D002478:Cells, Cultured; D005732:Gangliosides / Q000378:metabolism*; D008024:Ligands; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D011485:Protein Binding; D013744:Tetanus Toxin / Q000378:metabolism*
D005732:Gangliosides; D008024:Ligands; D013744:Tetanus Toxin