Literature summary for 3.4.24.69 extracted from

Ahmed, S.A.; Olson, M.A.; Ludivico, M.L.; Gilsdorf, J.; Smith, L.A.
Identification of residues surrounding the active site of type A botulinum neurotoxin important for substrate recognition and catalytic activity (2008), Protein J., 27, 151-162.

Search for more literature for 3.4.24.69

Application

Application	Comment	Organism
medicine	botulinum neurotoxins BoNT/A-G are used as therapeutics in a variety of neuromuscular disorders of the skeletal, glandular, and smooth muscles and pain disorders	Clostridium botulinum
molecular biology	botulinum neurotoxins BoNT/A-G are widely used as laboratory research tools	Clostridium botulinum
additional information	botulinum neurotoxins BoNT/A-G are the most potent of all toxins and potential bioterrorism agents, they are also used in cosmetic applications	Clostridium botulinum

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Clostridium botulinum

Protein Variants

Protein Variants	Comment	Organism
D130A	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
D369N	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
E163L	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
E163Q	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
E170A	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
E256A	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme in addition of ZnCl2 but not in absence of it, the ratio of activity in absence or presence of exogenous ZnCl2 is altered compared to the wild-type enzyme	Clostridium botulinum
E54A	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
E63A	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
K165L	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
Q161A	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
Q66A	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
R230K	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
R230L	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
R362L	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum
Y365F	site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme	Clostridium botulinum
Y365N	site-directed mutagenesis, the mutant shows reduced activity and an altered ratio of activity in absence or presence of exogenous ZnCl2 compared to the wild-type enzyme	Clostridium botulinum

Inhibitors

Inhibitors	Comment	Organism	Structure
Zn2+	addition of exogenous ZnCl2 to the assay mixture reduces the activity of BoNT/Am activity ratio of wild-type and mutant enzymes in presence or absence of ZnCl2, overview	Clostridium botulinum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetics of mutant enzymes, overview	Clostridium botulinum
2.92	-	SNAP25	pH 7.4, 37°C, recombinant wild-type enzyme in absence of exogenous ZnCl2	Clostridium botulinum
4.81	-	SNAP25	pH 7.4, 37°C, recombinant wild-type enzyme in presence of exogenous ZnCl2	Clostridium botulinum

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	zinc endopeptidase, dependent on, addition of ZnCl2 to the assay mixture reduces the activity of BoNT/A, activity ratio of wild-type and mutant enzymes in presence or absence of ZnCl2, overview, BoNT/A LC undergoes autocatalytic degradation into two major fragments in the presence of exogenous zinc	Clostridium botulinum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
SNAP25 + H2O	Clostridium botulinum	zinc-endopeptidase activity of the N-terminal light chain of BoNT/A on synaptosome-associated protein-25 kDa of the SNARE complex	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Clostridium botulinum	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
proteolytic modification	BoNT/A LC undergoes autocatalytic degradation into two major fragments in the presence of exogenous zinc, autolysis of wild-type and mtant BoNT/As, overview	Clostridium botulinum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	identification of active site and surrounding residues involved in substrate recognition and catalysis of BoNT/A, overview	Clostridium botulinum	?	-	?
SNAP25 + H2O	zinc-endopeptidase activity of the N-terminal light chain of BoNT/A on synaptosome-associated protein-25 kDa of the SNARE complex	Clostridium botulinum	?	-	?

Synonyms

Synonyms	Comment	Organism
type A botulinum neurotoxin	-	Clostridium botulinum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Clostridium botulinum

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
38	41	thermal denaturation of wild-type and mutant BoNT/As, overview	Clostridium botulinum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.41	-	SNAP25	pH 7.4, 37°C, recombinant wild-type enzyme in absence of exogenous ZnCl2	Clostridium botulinum
0.41	-	SNAP25	pH 7.4, 37°C, recombinant wild-type enzyme in presence of exogenous ZnCl2	Clostridium botulinum
12.35	-	SNAP25	pH 7.4, 37°C, recombinant wild-type enzyme in absence of exogenous ZnCl2	Clostridium botulinum
12.39	-	SNAP25	pH 7.4, 37°C, recombinant wild-type enzyme in presence of exogenous ZnCl2	Clostridium botulinum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Clostridium botulinum

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)