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Literature summary for 3.4.24.69 extracted from

  • Poras, H.; Ouimet, T.; Orng, S.V.; Fournie-Zaluski, M.C.; Popoff, M.R.; Roques, B.P.
    Detection and quantification of botulinum neurotoxin type A by a novel rapid in vitro fluorimetric assay (2009), Appl. Environ. Microbiol., 75, 4382-4390.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.24.69

Application

Application Comment Organism
medicine BoNT/A is widely used as a therapeutic agent and to reduce wrinkles. The toxin is used at very low doses, which have to be accurately quantified Clostridium botulinum
additional information botulinum neurotoxin type A is the most poisonous substance known to humans and is a potential bioterrorism agent Clostridium botulinum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00013
-
 Ac-IIGNLRH(Nle)ALD(Nle)GNEIDTQNRQIDRI(Nle)EKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum
0.00079
-
 Ac-IIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum
0.011
-
 Ac-KSDSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)LGSG-NH2 pH 7.4, 37°C Clostridium botulinum
0.013
-
 Ac-SNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+
-
 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. 25-kDa synaptosome-associated protein, is involved in acetylcholine release at the neuromuscular junction ?
-
 ?

Organism

Organism UniProt Comment Textmining
Clostridium botulinum
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ac-IIGNLRH(Nle)ALD(Nle)GNEIDTQNRQIDRI(Nle)EKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 + H2O i.e. peptide PL51, a SNAP-25-NH2in which all methionines were replaced by nonoxidizable Nle Clostridium botulinum Ac-IIGNLRH(Nle)ALD(Nle)GNEIDTQNRQIDRI(Nle)EKADSNKTRIDEAN(pNO2-Phe) + RA(1-pyrenylalanine)K(Nle)L-NH2
-
 ?
Ac-IIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 + H2O i.e. peptide PL50, a SNAP-25-NH2 acetylated at positions 156 to 203 [(pNO2-Phe)197, (1-pyrenylalanine)200, Nle202] Clostridium botulinum Ac-IIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEAN(pNO2-Phe) + RA(1-pyrenylalanine)K(Nle)L-NH2
-
 ?
Ac-KSDSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)LGSG-NH2 + H2O
-
 Clostridium botulinum Ac-KSDSNKTRIDEAN(pNO2-Phe) + RA(1-pyrenylalanine)K(Nle)LGSG-NH2
-
 ?
Ac-SNKTRIDEAN(1-pyrenylalanine)RA(pNO2-Phe)K(Nle)L-NH2 + H2O
-
 Clostridium botulinum Ac-SNKTRIDEAN(1-pyrenylalanine) + RA(pNO2-Phe)K(Nle)L-NH2
-
 ?
Ac-SNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 + H2O
-
 Clostridium botulinum Ac-SNKTRIDEAN(pNO2-Phe) + RA(1-pyrenylalanine)K(Nle)L-NH2
-
 ?
Ac-SNKTRIDEANQRATK(Nle)L-NH2 + H2O
-
 Clostridium botulinum Ac-SNKTRIDEANQ + RATK(Nle)L-NH2
-
 ?
additional information development of internally quenched fluorescent substrates containing the fluorophore/repressor pair pyrenylalanine/4-nitrophenylalanine for a sensitive assay method. (pNO2-Phe) and (1-pyrenylalanine) are, respectively, introduced at positions 197 and 200 of the cleavable fragment, amino acids 187 to 203, of SNAP-25, with norleucine at position 202 [Nle202], which is acetylated at its N terminus and amidated at its C-terminus. Sensitivity is increased when the peptide sequence of the previous substrate is lengthened to account for exosite binding to BoNT/A, substrate specificity and assay optimization, overview Clostridium botulinum ?
-
 ?
SNAP-25 + H2O i.e. 25-kDa synaptosome-associated protein Clostridium botulinum ?
-
 ?
SNAP-25 + H2O i.e. 25-kDa synaptosome-associated protein, is involved in acetylcholine release at the neuromuscular junction Clostridium botulinum ?
-
 ?
SNKTRIDEANQRATKML + H2O
-
 Clostridium botulinum SNKTRIDEANQ + RATKML
-
 ?

Synonyms

Synonyms Comment Organism
BoNT/A
-
 Clostridium botulinum
botulinum neurotoxin type A
-
 Clostridium botulinum

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.79
-
 Ac-SNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum
1.01
-
 Ac-KSDSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)LGSG-NH2 pH 7.4, 37°C Clostridium botulinum
1.15
-
 Ac-IIGNLRH(Nle)ALD(Nle)GNEIDTQNRQIDRI(Nle)EKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum
2.08
-
 Ac-IIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum

pH Optimum

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
61
-
 Ac-SNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum
92
-
 Ac-KSDSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)LGSG-NH2 pH 7.4, 37°C Clostridium botulinum
2630
-
 Ac-IIGNLRHMALDMGNEIDTQNRQIDRIMEKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum
8850
-
 Ac-IIGNLRH(Nle)ALD(Nle)GNEIDTQNRQIDRI(Nle)EKADSNKTRIDEAN(pNO2-Phe)RA(1-pyrenylalanine)K(Nle)L-NH2 pH 7.4, 37°C Clostridium botulinum