Literature summary for 3.4.24.69 extracted from

Kumaran, D.; Rawat, R.; Ludivico, M.L.; Ahmed, S.A.; Swaminathan, S.
Structure and substrate based inhibitor design for clostridium botulinum neurotoxin serotype A (2008), J. Biol. Chem., 283, 18883-18891.

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Application

Application	Comment	Organism
drug development	the enzyme is a target for design of specific inhibitors	Clostridium botulinum
additional information	the enzyme is used as a biowapon	Clostridium botulinum

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme free or in complex with substrate analogue inhibitor peptides, sitting drop vapor diffusion method at room temperature, 0.002 ml of 20 mg/ml enzyme in 2 mM DTT, 200 mM NaCl, and 20 mM HEPES, pH 7.4, is mixed with 0.002 ml reservoir solution containing 15% w/v PEG 3350, 0.3 M ammonium sulfate and 100 mM Bis-Tris buffer, pH 6.8 , equilibration against 0.8 ml of reservoir solution, plate-like crystals within a week, recombinant Balc424 is co-crystallized individually with the tetrapeptides, sitting drop vapor diffusion method at room temperature using conditions similar to native protein, Balc424 gives good complex crystals with RRGC, RRGM, RRGL and RRGI at stoichiometric ratios of 1:30, 1:30, 1:40 and 1:40, respectively, X-ray diffraction structure determination and analysis at 1.6-1.8 A resolution	Clostridium botulinum

Crystallization (Comment)

Organism

purified recombinant enzyme free or in complex with substrate analogue inhibitor peptides, sitting drop vapor diffusion method at room temperature, 0.002 ml of 20 mg/ml enzyme in 2 mM DTT, 200 mM NaCl, and 20 mM HEPES, pH 7.4, is mixed with 0.002 ml reservoir solution containing 15% w/v PEG 3350, 0.3 M ammonium sulfate and 100 mM Bis-Tris buffer, pH 6.8 , equilibration against 0.8 ml of reservoir solution, plate-like crystals within a week, recombinant Balc424 is co-crystallized individually with the tetrapeptides, sitting drop vapor diffusion method at room temperature using conditions similar to native protein, Balc424 gives good complex crystals with RRGC, RRGM, RRGL and RRGI at stoichiometric ratios of 1:30, 1:30, 1:40 and 1:40, respectively, X-ray diffraction structure determination and analysis at 1.6-1.8 A resolution

Clostridium botulinum

Inhibitors

Inhibitors	Comment	Organism
RRGC	an inhibitory substrate analogue tetrapeptide, binding structure, overview	Clostridium botulinum
RRGI	an inhibitory substrate analogue tetrapeptide, binding structure, overview	Clostridium botulinum
RRGL	an inhibitory substrate analogue tetrapeptide, binding structure, overview	Clostridium botulinum
RRGM	an inhibitory substrate analogue tetrapeptide, binding structure, overview	Clostridium botulinum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Clostridium botulinum	the seven antigenically distinct serotypes of Clostridium botulinum neurotoxins cleave specific SNARE complex proteins and block the release of neurotransmitters causing flaccid paralysis and are considered potential bioweapons, botulinum neurotoxin type A is the most potent among the clostridial neurotoxins	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Clostridium botulinum	P10845	seven antigenically distinct serotypes of neurotoxins	-

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	enzyme-substrate interaction at the active site, Tyr366 and Arg363 are important for catalysis, the S1' site is formed by Phe194, Thr215, Thr220, Asp370, and Arg363, reaction mechanism, overview	Clostridium botulinum	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the seven antigenically distinct serotypes of Clostridium botulinum neurotoxins cleave specific SNARE complex proteins and block the release of neurotransmitters causing flaccid paralysis and are considered potential bioweapons, botulinum neurotoxin type A is the most potent among the clostridial neurotoxins	Clostridium botulinum	?	-	?

Synonyms

Synonyms	Comment	Organism
Balc424	-	Clostridium botulinum
botulinum neurotoxin type A	-	Clostridium botulinum
Clostridium botulinum neurotoxin serotype A	-	Clostridium botulinum