title: Reductive methylation of lysine residues of botulinum neurotoxin types A and B.
abstract: Reductive methylation of botulinum neurotoxin (NT) serotypes A and B at various ratios of protein to reagent modified up to 75% of the lysine residues. Amino acid analysis of the modified proteins (HCl hydrolysed) confirmed selective modifications of lysine. The derivative N,N-dimethyl lysine was more abundant than monomethyl lysine; trimethyl lysine was not detected. Distribution of modified lysine residues among the heavy and light chains (Mr approximately 100,000 and approximately 50,000, respectively) of the dichain type A NT (Mr approximately 150,000) was approximately proportional to the lysine contents of the two subunit chains of the NT. Toxicity (mouse lethality) and serological reactivity (polyclonal antibody) of serotype A NT were not (or insignificantly) damaged following methylation of up to 72 lysine residues. Modification of 3 additional residues caused precipitous loss in toxicity. Toxicity of serotype B NT, unlike type A, appeared more sensitive to lysine modification. The large number of lysine residues that can be methylated without damaging toxicity of type A NT can be exploited to a) radiolabel the dichain protein exclusively in one chain keeping the other chain unlabelled, b) restrict the number of tryptic cleavage sites of the NT, and c) tag the protein with various markers or reactive ligands.
authors.affiliation: Food Research Institute, University of Wisconsin, Madison 53706.
authors.forename: V
authors.initials: V
authors.lastname: Sathyamoorthy
authors.forename: B R
authors.initials: BR
authors.lastname: DasGupta
mesh_terms: D000818:Animals; D001905:Botulinum Toxins / Q000633:toxicity*; D005779:Immunodiffusion; D007928:Lethal Dose 50; D008239:Lysine; D008745:Methylation; D051379:Mice; D008970:Molecular Weight; D009498:Neurotoxins; D010084:Oxidation-Reduction; D012703:Serotyping; D013329:Structure-Activity Relationship
chemical_list: D009498:Neurotoxins; D001905:Botulinum Toxins; D008239:Lysine