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Literature summary for 2.7.1.180 extracted from

  • Fang, X.; Liang, P.; Raba, D.; Rosas-Lemus, M.; Chakravarthy, S.; Tuz, K.; Juárez, O.
    Kinetic characterization of Vibrio cholerae ApbE substrate specificity and regulatory mechanisms (2017), PLoS ONE, 12, e0186805 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.1.180

Activating Compound

Activating Compound Comment Organism Structure
ADP ADP does not inhibit the transfer reaction, but significantly activates it by 6fold Vibrio cholerae

Cloned(Commentary)

Cloned (Comment) Organism
gene apbE, recombinant overexpression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli. The recombinant protein does not contain the leader sequence (first 17 amino acids), and it is obtained as a cytosolic protein, although it is naturally localized in the periplasmic space Vibrio cholerae

Protein Variants

Protein Variants Comment Organism
H257G site-directed mutagenesis, the mutant's flavin transfer activity is abolished. After reconstitution, the H257G shows a FAD:protein ratio nearly identical to the wild-type Vibrio cholerae

Inhibitors

Inhibitors Comment Organism Structure
AMP 1 mM of AMP has no effect on the activity, but a higher concentration (10 mM), produces a slight inhibition Vibrio cholerae
ATP 1 mM of ATP has no effect on the activity, but a higher concentration (10 mM), produces a slight inhibition Vibrio cholerae
Ba2+
-
 Vibrio cholerae
Ca2+
-
 Vibrio cholerae
EDTA
-
 Vibrio cholerae
FMN 1 mM of FMN has no effect on the activity, but a higher concentration (10 mM), produces a slight inhibition Vibrio cholerae
n-dodecyl-beta-D-maltopyranoside 0.05% n-dodecyl-beta-D-maltopyranoside causes 20% inhibition of the enzyme Vibrio cholerae
Zn2+
-
 Vibrio cholerae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information ApbE activity is highly sensitive to FAD, and is saturated at 0.001 mM Vibrio cholerae
0.00006
-
 FAD pH 9.0, 22°C, recombinant wild-type enzyme Vibrio cholerae
0.1
-
 [Vibrio cholerae protein NqrC]-L-threonine above, pH 9.0, 22°C, recombinant wild-type enzyme Vibrio cholerae

Localization

Localization Comment Organism GeneOntology No. Textmining
periplasm
-
 Vibrio cholerae
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Cd2+ highly activates by 4fold Vibrio cholerae
Co2+ activates Vibrio cholerae
Mg2+ required for catalysis, Mg2+ does not play a major role in stabilizing the interaction of the protein with FAD, Mg2+ in the preferred divalent cation Vibrio cholerae
Mn2+ activates Vibrio cholerae
additional information divalent cations are essential for ApbE activity, and their removal by EDTA abolishes the activity. ApbE is also able to use other divalent cations, such as Mn2+ and Co2+, obtaining a similar activity compared to Mg2+. The coordination sphere of ApbE enzymes largely determines the specificity of the enzyme for the divalent cation Vibrio cholerae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37000
-
 recombinant enzyme, gel filtration Vibrio cholerae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
FAD + [protein]-L-threonine Vibrio cholerae
-
 [protein]-FMN-L-threonine + AMP
-
 ?
FAD + [Vibrio cholerae protein NqrC]-L-threonine Vibrio cholerae
-
 [Vibrio cholerae protein NqrC]-FMN-L-threonine + AMP
-
 ?

Organism

Organism UniProt Comment Textmining
Vibrio cholerae A0A0F4FI39
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type ApbE and mutants from Escherichia coli by ultrafiltration, nickel affinity chromatography, cation chromatography, and gel filtration, to homogeneity Vibrio cholerae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FAD + [protein]-L-threonine
-
 Vibrio cholerae [protein]-FMN-L-threonine + AMP
-
 ?
FAD + [Vibrio cholerae protein NqrC]-L-threonine
-
 Vibrio cholerae [Vibrio cholerae protein NqrC]-FMN-L-threonine + AMP
-
 ?
FAD + [Vibrio cholerae protein NqrC]-L-threonine conserved Lys207 in NqrC can account for the pH dependency Vibrio cholerae [Vibrio cholerae protein NqrC]-FMN-L-threonine + AMP
-
 ?
additional information substrate specificity, overview. FAD cannot be substituted by FMN Vibrio cholerae ?
-
-

Subunits

Subunits Comment Organism
monomer 1 * 37000, about, recombinant enzyme, SDS-PAGE Vibrio cholerae

Synonyms

Synonyms Comment Organism
apbE
-
 Vibrio cholerae
flavin transferase
-
 Vibrio cholerae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Vibrio cholerae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 9 broad optimum Vibrio cholerae

General Information

General Information Comment Organism
evolution ApbE is a member of a family of flavin transferases that incorporates flavin mononucleotide (FMN) to subunits of diverse respiratory complexes, which fulfill important homeostatic functions. Enzyme residue His257 is absolutely conserved in the family Vibrio cholerae
physiological function substrate specificity and regulatory mechanisms, overview. Residue H257 is the residue whose deprotonation controls the activity, it plays an important role in the catalytic mechanism of ApbE. Residue His257 is indeed essential for catalysis, but not for substrate binding Vibrio cholerae