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Mutations at a glycine loop in aminolevulinate synthase affect pyridoxal phosphate cofactor binding and catalysis

Gong, J.; Kay, C.J.; Barber, M.J.; Ferreira, G.C.; Biochemistry 35, 14109-14117 (1996)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.3.1.37
expression of wild-type and mutants in Escherichia coli
Mus musculus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.3.1.37
G142C
glycine-rich motif mutant, 15fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 6% turnover compared to the wild-type, 4fold increase of Km-value for glycine
Mus musculus
2.3.1.37
G144A
glycine-rich motif mutant, 8.5fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 43% turnover compared to the wild-type, unaltered Km-values for the substrates
Mus musculus
2.3.1.37
G144S
glycine-rich motif mutant, 8fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 39% turnover compared to the wild-type, unaltered Km-values for the substrates
Mus musculus
2.3.1.37
G144T
glycine-rich motif mutant, 24.5fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 21% turnover compared to the wild-type, unaltered Km-values for the substrates
Mus musculus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.1.37
0.00064
-
succinyl-CoA
recombinant mutant G144A
Mus musculus
2.3.1.37
0.0012
-
succinyl-CoA
recombinant erythroid isoform and mutant G144S
Mus musculus
2.3.1.37
0.0019
-
succinyl-CoA
recombinant mutant G144T
Mus musculus
2.3.1.37
0.002
-
succinyl-CoA
recombinant mutant G142C
Mus musculus
2.3.1.37
0.002
-
succinyl-CoA
recombinant mutant G142C
Rattus norvegicus
2.3.1.37
6.2
-
glycine
recombinant mutant G144T
Mus musculus
2.3.1.37
9.3
-
glycine
recombinant mutant G144A
Mus musculus
2.3.1.37
11.9
-
glycine
recombinant mutant G144S
Mus musculus
2.3.1.37
12.5
-
glycine
recombinant erythroid isoform
Mus musculus
2.3.1.37
52.2
-
glycine
recombinant mutant G142C
Mus musculus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.3.1.37
Mus musculus
-
-
-
2.3.1.37
Rattus norvegicus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.3.1.37
recombinant wild-type and mutants from E. coli
Mus musculus
Reaction
EC Number
Reaction
Commentary
Organism
2.3.1.37
succinyl-CoA + glycine = 5-aminolevulinate + CoA + CO2
pyridoxal 5'-phosphate binding site, sequence and function of glycine-rich motif; Tyr121, Asp279, Arg439, and Lys313 are involved in substrate and cofactor binding, mechanism, subunit localisation
Mus musculus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.37
succinyl-CoA + glycine
-
486851
Mus musculus
5-aminolevulinate + CoA + CO2
-
486851
Mus musculus
?
2.3.1.37
succinyl-CoA + glycine
-
486851
Rattus norvegicus
5-aminolevulinate + CoA + CO2
-
486851
Rattus norvegicus
?
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.3.1.37
additional information
-
thermal stability of glycine-rich motif mutants is reduced compared to the wild-type
Mus musculus
2.3.1.37
additional information
-
-
Rattus norvegicus
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.3.1.37
0.0183
-
succinyl-CoA
recombinant erythroid isoform mutant G142C
Mus musculus
2.3.1.37
0.0633
-
succinyl-CoA
recombinant erythroid isoform mutant G144T
Mus musculus
2.3.1.37
0.12
-
succinyl-CoA
recombinant erythroid isoform mutant G144S
Mus musculus
2.3.1.37
0.132
-
succinyl-CoA
recombinant erythroid isoform mutant G144A
Mus musculus
2.3.1.37
0.308
-
succinyl-CoA
recombinant erythroid isoform
Mus musculus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.3.1.37
pyridoxal 5'-phosphate
-
Rattus norvegicus
2.3.1.37
pyridoxal 5'-phosphate
-
Mus musculus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.37
expression of wild-type and mutants in Escherichia coli
Mus musculus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.3.1.37
pyridoxal 5'-phosphate
-
Rattus norvegicus
2.3.1.37
pyridoxal 5'-phosphate
-
Mus musculus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.3.1.37
G142C
glycine-rich motif mutant, 15fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 6% turnover compared to the wild-type, 4fold increase of Km-value for glycine
Mus musculus
2.3.1.37
G144A
glycine-rich motif mutant, 8.5fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 43% turnover compared to the wild-type, unaltered Km-values for the substrates
Mus musculus
2.3.1.37
G144S
glycine-rich motif mutant, 8fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 39% turnover compared to the wild-type, unaltered Km-values for the substrates
Mus musculus
2.3.1.37
G144T
glycine-rich motif mutant, 24.5fold increased dissociation constant value for binding of cofactor pyridoxal 5'-phosphate, 21% turnover compared to the wild-type, unaltered Km-values for the substrates
Mus musculus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.3.1.37
0.00064
-
succinyl-CoA
recombinant mutant G144A
Mus musculus
2.3.1.37
0.0012
-
succinyl-CoA
recombinant erythroid isoform and mutant G144S
Mus musculus
2.3.1.37
0.0019
-
succinyl-CoA
recombinant mutant G144T
Mus musculus
2.3.1.37
0.002
-
succinyl-CoA
recombinant mutant G142C
Mus musculus
2.3.1.37
0.002
-
succinyl-CoA
recombinant mutant G142C
Rattus norvegicus
2.3.1.37
6.2
-
glycine
recombinant mutant G144T
Mus musculus
2.3.1.37
9.3
-
glycine
recombinant mutant G144A
Mus musculus
2.3.1.37
11.9
-
glycine
recombinant mutant G144S
Mus musculus
2.3.1.37
12.5
-
glycine
recombinant erythroid isoform
Mus musculus
2.3.1.37
52.2
-
glycine
recombinant mutant G142C
Mus musculus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.37
recombinant wild-type and mutants from E. coli
Mus musculus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.3.1.37
succinyl-CoA + glycine
-
486851
Mus musculus
5-aminolevulinate + CoA + CO2
-
486851
Mus musculus
?
2.3.1.37
succinyl-CoA + glycine
-
486851
Rattus norvegicus
5-aminolevulinate + CoA + CO2
-
486851
Rattus norvegicus
?
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.3.1.37
additional information
-
thermal stability of glycine-rich motif mutants is reduced compared to the wild-type
Mus musculus
2.3.1.37
additional information
-
-
Rattus norvegicus
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.3.1.37
0.0183
-
succinyl-CoA
recombinant erythroid isoform mutant G142C
Mus musculus
2.3.1.37
0.0633
-
succinyl-CoA
recombinant erythroid isoform mutant G144T
Mus musculus
2.3.1.37
0.12
-
succinyl-CoA
recombinant erythroid isoform mutant G144S
Mus musculus
2.3.1.37
0.132
-
succinyl-CoA
recombinant erythroid isoform mutant G144A
Mus musculus
2.3.1.37
0.308
-
succinyl-CoA
recombinant erythroid isoform
Mus musculus