Literature summary for 3.4.24.69 extracted from

Silvaggi, N.R.; Boldt, G.E.; Hixon, M.S.; Kennedy, J.P.; Tzipori, S.; Janda, K.D.; Allen, K.N.
Structures of Clostridium botulinum neurotoxin serotype A light chain complexed with small-molecule inhibitors highlight active-site flexibility (2007), Chem. Biol., 14, 533-542.

Search for more literature for 3.4.24.69

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged truncated enzyme, residues 1-424, in Escherichia coli strain BL21(DE3)	Clostridium botulinum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged truncated enzyme, residues 1-424, in complex with inhibitors 4-chlorocinnamic hydroxamate, 2,4-dichlorocinnamic hydroxamate, and L-arginine hydroxamate, hanging drop vapor diffusion method, drops are formed of equal parts BoNT/A-LC(1-424) at 10 mg/ml in 20 mM HEPES, pH 7.5, 50 mM NaCl and well solution containing 10%-15% PEG-2000 monomethyl ether, 0.3M(NH4)2HPO4, 50 mM Tris, pH 8.5, co-crystallization with inhibitor by addition of 0.5 mM ligand to the protein solution, 1.3 days, larger crystals by microseeding, X-ray diffraction structure determination and analysis at 1.9-2.5 A resolution, modeling	Clostridium botulinum

Crystallization (Comment)

Organism

purified recombinant His-tagged truncated enzyme, residues 1-424, in complex with inhibitors 4-chlorocinnamic hydroxamate, 2,4-dichlorocinnamic hydroxamate, and L-arginine hydroxamate, hanging drop vapor diffusion method, drops are formed of equal parts BoNT/A-LC(1-424) at 10 mg/ml in 20 mM HEPES, pH 7.5, 50 mM NaCl and well solution containing 10%-15% PEG-2000 monomethyl ether, 0.3M(NH4)2HPO4, 50 mM Tris, pH 8.5, co-crystallization with inhibitor by addition of 0.5 mM ligand to the protein solution, 1.3 days, larger crystals by microseeding, X-ray diffraction structure determination and analysis at 1.9-2.5 A resolution, modeling

Clostridium botulinum

Inhibitors

Inhibitors	Comment	Organism
2,4-dichlorocinnamic hydroxamate	binding site and complex structure, overview	Clostridium botulinum
4-chlorocinnamic hydroxamate	binding site and complex structure, overview	Clostridium botulinum
L-Arginine hydroxamate	binding site and complex structure, overview	Clostridium botulinum

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	binding site structure, a long a helix contains the HEXXH Zn(II)-binding motif, modeling, overview	Clostridium botulinum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
SNAP-25 + H2O	Clostridium botulinum	i.e. synaptosomal associated protein of 25 kDa	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Clostridium botulinum	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged truncated enzyme, residues 1-424, from Escherichia coli strain BL21(DE3) by metal ion affinity chromatography	Clostridium botulinum

Reaction

Reaction	Comment	Organism	Reaction ID
Limited hydrolysis of proteins of the neuroexocytosis apparatus, synaptobrevin (also known as neuronal vesicle-associated membrane protein, VAMP), synaptosome-associated protein of 25 kDa (SNAP25) or syntaxin. No detected action on small molecule substrates	active site structure and structure-function relationship	Clostridium botulinum	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
SNAP-25 + H2O	i.e. synaptosomal associated protein of 25 kDa	Clostridium botulinum	?	-	?
SNAP-25 + H2O	i.e. synaptosomal associated protein of 25 kDa, all botulinus neurotoxin serotypes cleave the substrate at a unique peptide bond, BoNT/A cleaves SNAP-25 between residues Gln197 and Arg198. Phe194, Ile161, and Asp370 form the S1' subsite responsible for binding the P1' arginine side chain of SNAP-25, overview	Clostridium botulinum	?	-	?

Synonyms

Synonyms	Comment	Organism
Clostridium botulinum neurotoxin serotype A light chain	-	Clostridium botulinum

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.0003	-	-	Clostridium botulinum	2,4-dichlorocinnamic hydroxamate
0.015	-	-	Clostridium botulinum	4-chlorocinnamic hydroxamate
0.06	-	-	Clostridium botulinum	L-Arginine hydroxamate