Crystal structure of the catalytic domain of botulinum neurotoxin subtype A3

Abstract

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How Botulinum Neurotoxin Light Chain A1 Maintains Stable Association with the Intracellular Neuronal Plasma Membrane

“…The current study sought to establish how the previously identified LHD (268-357) mediated LC/A1 association with the plasma membrane [35]. A co-crystal of LC/A1 bound to SNAP-25 (residues 146-204 (PDB: 1XTG)) and the recently solved crystal structure of LC/A3 [43] showed the LHD to be organized into three sequential structural regions (R1-R3 (Figure 1). R1, residues 275-300, included an α-helix; R2, residues 302-334, included a loop, α-helix, and loop; and R3, residues 335-357, included an α-helix, loop, and α-helix.…”

A DARPin promotes faster onset of botulinum neurotoxin A1 action

“…Protein expression and purification were performed as described previously 66 . Proteins were expressed in E. coli strain BL21(DE3) (NEB).…”

Resolution of Two Steps in Botulinum Neurotoxin Serotype A1 Light Chain Localization to the Intracellular Plasma Membrane

“…Within N, A1 and A2 were identical, while A3LM possessed a unique K 11 R substitution. Within the LHD, A1 and A2 were 97% identical, while A3LM had ~60% primary amino acid homology with A1 and A2, with differences in surface charge potential as there is a cluster of basic amino acids present in A3LM that are absent in A1 [31]. Supporting roles for the N and LHD in intracellular LC targeting, such as A1 ectopically expressing A2 at steady state, were localized to the plasma membrane (Supplemental Figure S1, see Supplementary Materials). )…”

“…Supporting roles for the N and LHD in intracellular LC targeting, such as A1 ectopically expressing A2 at steady state, were localized to the plasma membrane (Supplemental Figure S1, see Supplementary Materials). ) and middle A3LM (PDB: 7DVL) [31] and right A1 and A3LM merged with N (residues 1-17, red) and LHD (268-357, green) highlighted. Merge of A1 and A3LM was prepared with PyMol software.…”

“…(B) Structural alignments of A1 and A3LM. The light chain (LC) is shown in gray, the N in red, and LHD in green A1 (PDB: 1XTG) and middle A3LM (PDB: 7DVL)[31] and right A1 and A3LM merged with N (residues 1-17, red) and LHD (268-357, green) highlighted. Merge of A1 and A3LM was prepared with PyMol software.…”