title: Identification of novel small molecule ligands that bind to tetanus toxin.
abstract: Tetanus toxin belongs to a family of clostridial protein neurotoxins for which there are no known antidotes. Another closely related member of this family, botulinum toxin, is being used with increasing frequency by physicians to treat severe muscle disorders. Botulinum toxin has also been produced in large quantities by terrorists for use as a biological weapon. To identify small molecule ligands that might bind to the targeting domain of tetanus and botulinum toxins and to facilitate the design of inhibitors and new reagents for their detection, molecular docking calculations were used to screen a large database of compounds for their potential to bind to the C fragment of tetanus toxin. Eleven of the predicted ligands were assayed by electrospray ionization mass spectrometry (ESI-MS) for binding to the tetanus toxin C fragment, and five ligands (45%) were found to bind to the protein. One of these compounds, doxorubicin, was observed to have strong hydrophobic interactions with the C fragment. To check the ligands for their ability to compete with ganglioside binding, each was also tested using a GT1b liposome assay. Doxorubicin was the only ligand found to competitively bind the tetanus toxin C fragment with an appreciable binding constant (9.4 microM).
authors.affiliation: Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94550, USA.
authors.forename: F C
authors.initials: FC
authors.lastname: Lightstone
authors.forename: M C
authors.initials: MC
authors.lastname: Prieto
authors.forename: A K
authors.initials: AK
authors.lastname: Singh
authors.forename: M C
authors.initials: MC
authors.lastname: Piqueras
authors.forename: R M
authors.initials: RM
authors.lastname: Whittal
authors.forename: M S
authors.initials: MS
authors.lastname: Knapp
authors.forename: R
authors.initials: R
authors.lastname: Balhorn
authors.forename: D C
authors.initials: DC
authors.lastname: Roe
mesh_terms: D001665:Binding Sites; D003198:Computer Simulation; D004317:Doxorubicin / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D008081:Liposomes / Q000378:metabolism; D013058:Mass Spectrometry; D008958:Models, Molecular; D008968:Molecular Conformation; D010446:Peptide Fragments / Q000737:chemistry; D013744:Tetanus Toxin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008024:Ligands; D008081:Liposomes; D010446:Peptide Fragments; D013744:Tetanus Toxin; D004317:Doxorubicin