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Sirt5 deacylation activities show differential sensitivities to nicotinamide inhibition

Fischer, F.; Gertz, M.; Suenkel, B.; Lakshminarasimhan, M.; Schutkowski, M.; Steegborn, C.; PLoS One 7, e45098 (2012)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.3.1.B41
-
Archaeoglobus fulgidus
2.3.1.B43
-
Homo sapiens
2.3.1.286
-
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.3.1.B41
M70R
further decrease in nicotinamide sensitivity compared to wild-type
Archaeoglobus fulgidus
2.3.1.B43
R105L
mutant retains wild-type deacetylation activity and partially gains Sirt3-like nicotinamide sensitivity (IC50: 0.189 mM)
Homo sapiens
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.1.B41
additional information
nicotinamide insensitivity, IC50: 2.1 mM
Archaeoglobus fulgidus
2.3.1.B43
nicotinamide
nicotinamide inhibition of Sirt5-dependent deacetylation features weak substrate-dependence but requires high, nonphysiological nicotinamide concentrations. The insensitivity of Sirt5 to physiological nicotinamide concentrations is due to a lack of non-competitive inhibition via base exchange, and the inhibition observed at very high nicotinamide concentrations is likely caused by competition with NAD+
Homo sapiens
2.3.1.286
nicotinamide
0.5 mM, complete inhibition
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.1.B41
0.22
-
SKEYFS(N6-acetyl)KQK
pH 7.8, 37°C
Archaeoglobus fulgidus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.3.1.B41
Archaeoglobus fulgidus
O30124
-
-
2.3.1.B43
Homo sapiens
Q9NXA8
-
-
2.3.1.286
Homo sapiens
Q9NTG7
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.3.1.B41
-
Archaeoglobus fulgidus
2.3.1.B43
-
Homo sapiens
2.3.1.286
-
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.B41
additional information
insignificant activity with SKEYFS(N6-succinyl)KQK
730706
Archaeoglobus fulgidus
?
-
-
-
-
2.3.1.B41
NAD+ + SKEYFS(N6-acetyl)KQK
-
730706
Archaeoglobus fulgidus
nicotinamide + SKEYFSKQK + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.B43
NAD+ + benzoyl-RGVL(N6-succinyl)KEYGV-amide
carbamoyl phosphate synthetase 1 Lys527 peptide
730706
Homo sapiens
nicotinamide + benzoyl-RGVLKEYGV-amide + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.B43
NAD+ + FKRGVL(N6-acetyl)KEYGVKV
carbamoyl phosphate synthetase 1 Lys527 peptide
730706
Homo sapiens
nicotinamide + FKRGVLKEYGVKV + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.B43
NAD+ + TRSG(N6-acetyl)KVMR
peptide based on an acetyl-CoA synthetase 2 acetylation site
730706
Homo sapiens
nicotinamide + TRSGKVMR + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.286
NAD+ + TRSG(N6-acetyl)KVMR
peptide based on an acetyl-CoA synthetase 2 acetylation site
730706
Homo sapiens
nicotinamide + TRSGKVMR + 2'-O-acetyl-ADP-ribose
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.1.B41
37
-
assay at
Archaeoglobus fulgidus
2.3.1.B43
37
-
assay at
Homo sapiens
2.3.1.286
37
-
assay at
Homo sapiens
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.B41
7.8
-
assay at
Archaeoglobus fulgidus
2.3.1.B43
7.8
-
assay at
Homo sapiens
2.3.1.286
7.8
-
assay at
Homo sapiens
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
2.3.1.B43
0.021
-
pH 7.8, 37°C, substrate: benzoyl-RGVL(succK)EYGV-amide
Homo sapiens
nicotinamide
2.3.1.B43
0.7
-
pH 7.8, 37°C, substrate: TRSG(acK)VMR
Homo sapiens
nicotinamide
2.3.1.B43
1.6
-
pH 7.8, 37°C, substrate: FKRGVL(acK)EYGVKV
Homo sapiens
nicotinamide
2.3.1.286
0.043
-
pH 7.8, 37°C, substrate: TRSG(N6-acetyl)KVMR
Homo sapiens
nicotinamide
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.B41
-
Archaeoglobus fulgidus
2.3.1.B43
-
Homo sapiens
2.3.1.286
-
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.3.1.B41
M70R
further decrease in nicotinamide sensitivity compared to wild-type
Archaeoglobus fulgidus
2.3.1.B43
R105L
mutant retains wild-type deacetylation activity and partially gains Sirt3-like nicotinamide sensitivity (IC50: 0.189 mM)
Homo sapiens
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
2.3.1.B43
0.021
-
pH 7.8, 37°C, substrate: benzoyl-RGVL(succK)EYGV-amide
Homo sapiens
nicotinamide
2.3.1.B43
0.7
-
pH 7.8, 37°C, substrate: TRSG(acK)VMR
Homo sapiens
nicotinamide
2.3.1.B43
1.6
-
pH 7.8, 37°C, substrate: FKRGVL(acK)EYGVKV
Homo sapiens
nicotinamide
2.3.1.286
0.043
-
pH 7.8, 37°C, substrate: TRSG(N6-acetyl)KVMR
Homo sapiens
nicotinamide
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.3.1.B41
additional information
nicotinamide insensitivity, IC50: 2.1 mM
Archaeoglobus fulgidus
2.3.1.B43
nicotinamide
nicotinamide inhibition of Sirt5-dependent deacetylation features weak substrate-dependence but requires high, nonphysiological nicotinamide concentrations. The insensitivity of Sirt5 to physiological nicotinamide concentrations is due to a lack of non-competitive inhibition via base exchange, and the inhibition observed at very high nicotinamide concentrations is likely caused by competition with NAD+
Homo sapiens
2.3.1.286
nicotinamide
0.5 mM, complete inhibition
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.1.B41
0.22
-
SKEYFS(N6-acetyl)KQK
pH 7.8, 37°C
Archaeoglobus fulgidus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.B41
-
Archaeoglobus fulgidus
2.3.1.B43
-
Homo sapiens
2.3.1.286
-
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.B41
additional information
insignificant activity with SKEYFS(N6-succinyl)KQK
730706
Archaeoglobus fulgidus
?
-
-
-
-
2.3.1.B41
NAD+ + SKEYFS(N6-acetyl)KQK
-
730706
Archaeoglobus fulgidus
nicotinamide + SKEYFSKQK + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.B43
NAD+ + benzoyl-RGVL(N6-succinyl)KEYGV-amide
carbamoyl phosphate synthetase 1 Lys527 peptide
730706
Homo sapiens
nicotinamide + benzoyl-RGVLKEYGV-amide + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.B43
NAD+ + FKRGVL(N6-acetyl)KEYGVKV
carbamoyl phosphate synthetase 1 Lys527 peptide
730706
Homo sapiens
nicotinamide + FKRGVLKEYGVKV + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.B43
NAD+ + TRSG(N6-acetyl)KVMR
peptide based on an acetyl-CoA synthetase 2 acetylation site
730706
Homo sapiens
nicotinamide + TRSGKVMR + 2'-O-acetyl-ADP-ribose
-
-
-
?
2.3.1.286
NAD+ + TRSG(N6-acetyl)KVMR
peptide based on an acetyl-CoA synthetase 2 acetylation site
730706
Homo sapiens
nicotinamide + TRSGKVMR + 2'-O-acetyl-ADP-ribose
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.3.1.B41
37
-
assay at
Archaeoglobus fulgidus
2.3.1.B43
37
-
assay at
Homo sapiens
2.3.1.286
37
-
assay at
Homo sapiens
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.3.1.B41
7.8
-
assay at
Archaeoglobus fulgidus
2.3.1.B43
7.8
-
assay at
Homo sapiens
2.3.1.286
7.8
-
assay at
Homo sapiens
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.3.1.B41
0.132
-
SKEYFS(N6-acetyl)KQK
pH 7.8, 37°C
Archaeoglobus fulgidus
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.3.1.B41
0.132
-
SKEYFS(N6-acetyl)KQK
pH 7.8, 37°C
Archaeoglobus fulgidus