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Literature summary for 3.4.24.69 extracted from

  • Salzameda, N.T.; Barbieri, J.T.; Janda, K.D.
    Synthetic substrate for application in both high and low throughput assays for botulinum neurotoxin B protease inhibitors (2009), Bioorg. Med. Chem. Lett., 19, 5848-5850.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.24.69

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0016
-
 VAMP2 pH 7.4, 22°C Clostridium botulinum
0.0709
-
 Ac-ERDQKLSELDDRADALQAG-(7-methoxy-4-methylcoumaryl)Lys-SQ-diaminopropionic acid(2,4-dinitrophenyl)-ESSAAKLKRKYWWKNLK-NH2 pH 7.4, 22°C Clostridium botulinum

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ BoNT/B is a zinc endopeptidase Clostridium botulinum

Organism

Organism UniProt Comment Textmining
Clostridium botulinum
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Ac-ERDQKLSELDDRADALQAG-(7-methoxy-4-methylcoumaryl)Lys-SQ-diaminopropionic acid(2,4-dinitrophenyl)-ESSAAKLKRKYWWKNLK-NH2 + H2O development of a FRET peptide substrate, based on the native substrate binding site of human VAMP2 residues 55-94, and evaluation for enzymatic cleavage by the BoNT/B light chain protease, overview. For the synthesis position 74 is mutated to Lys in order to couple 7-methoxycoumarin-4-acetic acid, MCA, to the amine via an amide bond, in part to aid in the flexibility of the MCA to allow free rotation away from the active site and not affect binding and/or cleavage of the peptide. At position 77 the native Phe is replaced with the unnatural amino acid diaminopropionic acid to facilitate coupling of 2,4-dinitrophenyl to the peptide. Thr79 is mutated to a serine increasing kcat 2fold without affecting Km Clostridium botulinum ?
-
 ?
VAMP2 + H2O human VAMP2 substrate, i.e. vesicle-associated membrane protein 2 Clostridium botulinum ?
-
 ?

Synonyms

Synonyms Comment Organism
BoNT/B
-
 Clostridium botulinum
BoNT/B light chain protease
-
 Clostridium botulinum
botulinum neurotoxin B protease
-
 Clostridium botulinum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Clostridium botulinum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1
-
 VAMP2 pH 7.4, 22°C Clostridium botulinum
3.81
-
 Ac-ERDQKLSELDDRADALQAG-(7-methoxy-4-methylcoumaryl)Lys-SQ-diaminopropionic acid(2,4-dinitrophenyl)-ESSAAKLKRKYWWKNLK-NH2 pH 7.4, 22°C Clostridium botulinum

pH Optimum

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