Genetic analysis of bacterial acetyltransferases: identification of amino acids determining the specificities of the aminoglycoside 6-N-acetyltransferase Ib and IIa proteins
Rather, P.N.; Munayyer, H.; Mann, P.A.; Hare, R.S.; Miller, G.H.; Shaw, K.J.; J. Bacteriol. 174, 3196-3203 (1992) 
Data extracted from this reference:
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
2.3.1.82
isogenic plasmids pSCH4663 containing aac(6')-Ib gene and pSCHB4105 containing aac(6')-IIa gene, transferred to Escherichia coli DH5alpha
Escherichia coli
Engineering
EC Number
Protein Variants
Commentary
Organism
2.3.1.82
L119S
AAC(6')-Ib L119S. The AAC(6')-Ib protein is unable to efficiently modify gentamicin C1, 1.7% relative to sisomicin, however it is capable of modifying amikacin, 65.5% relative to sisomycin. The mutation results in a 2.8fold increase in acetylation of gentamicin C1, but causes an 8.7fold reduction in the ability to modify amikacin
Escherichia coli
2.3.1.82
S119L
AAC(6')-IIa S119L. The AAC(6')-IIa protein modifies gentamicin C1 at 10.1% relative to sisomicin, however it shows low activity towards amikacin, 4.1% relative to sisomycin. The mutation results in a 4.8fold reduction in the acetylation of gentamicin C1, but causes an 2fold increase in the ability to modify amikacin
Escherichia coli
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2.3.1.82
additional information
Escherichia coli
involved in resistance to antibiotics
?
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
2.3.1.82
Escherichia coli
-
DH5alpha, AAC(6')-Ib protein and AAC(6')-IIa protein
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2.3.1.82
acetyl-CoA + 2'-N-ethylnetilmicin
-
636903
Escherichia coli
CoA + N6'-acetyl-2'-N-ethylnetilmicin
-
-
-
?
2.3.1.82
acetyl-CoA + amikacin
the AAC(6')-Ib protein is unable to efficiently modify gentamicin C1, 1.7% relative activity to sisomicin, however it is capable of modifying amikacin, 65.5% relative activity to sisomycin. The mutant enzyme AAC(6')-Ib L119S shows a 2.8fold increase in acetylation of gentamicin C1, but a 8.7fold reduction in the ability to modify amikacin. The AAC(6')-IIa protein modifies gentamicin C1 at 10.1% relative activity to sisomicin, however it shows low activity towards amikacin, 4.1% activity relative to sisomycin. The mutation AAC(6')-IIa S119L results in a 4.8fold reduction in the acetylation of gentamicin C1, but causes an 2fold increase in the ability to modify amikacin
636903
Escherichia coli
CoA + N6'-acetylamikacin
-
-
-
?
2.3.1.82
acetyl-CoA + gentamicin C1
the AAC(6')-Ib protein is unable to efficiently modify gentamicin C1, 1.7% relative activity to sisomicin, however it is capable of modifying amikacin, 65.5% relative activity to sisomycin. The mutant enzyme AAC(6')-Ib L119S shows a 2.8fold increase in acetylation of gentamicin C1, but a 8.7fold reduction in the ability to modify amikacin. The AAC(6')-IIa protein modifies gentamicin C1 at 10.1% relative activity to sisomicin, however it shows low activity towards amikacin, 4.1% activity relative to sisomycin. The mutation AAC(6')-IIa S119L results in a 4.8fold reduction in the acetylation of gentamicin C1, but causes an 2fold increase in the ability to modify amikacin
636903
Escherichia coli
CoA + N6'-acetylgentamicin C1
-
-
-
?
2.3.1.82
acetyl-CoA + gentamicin C1a
poor substrate, AAC-(6')-II type
636903
Escherichia coli
CoA + N6'-acetylgentamicin C1a
-
-
-
?
2.3.1.82
acetyl-CoA + sisomicin
-
636903
Escherichia coli
CoA + N6'-acetylsisomicin
-
-
-
?
2.3.1.82
acetyl-CoA + tobramycin
-
636903
Escherichia coli
CoA + N6'-acetyltobramycin
-
-
-
?
2.3.1.82
additional information
a single amino acid, Leu119 in AAC(6')-Ib and S119 in AAC(6')-IIa, is largely responsible for determining the specificity of the AAC(6')-Ib and AAC(6')-IIa proteins. Changing this amino acid in either the AAC(6')-Ib or the AAC(6')-IIa protein results in a dramatic change in substrate specificity
636903
Escherichia coli
?
-
-
-
?
2.3.1.82
additional information
involved in resistance to antibiotics
636903
Escherichia coli
?
-
-
-
?
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.3.1.82
isogenic plasmids pSCH4663 containing aac(6')-Ib gene and pSCHB4105 containing aac(6')-IIa gene, transferred to Escherichia coli DH5alpha
Escherichia coli
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
2.3.1.82
L119S
AAC(6')-Ib L119S. The AAC(6')-Ib protein is unable to efficiently modify gentamicin C1, 1.7% relative to sisomicin, however it is capable of modifying amikacin, 65.5% relative to sisomycin. The mutation results in a 2.8fold increase in acetylation of gentamicin C1, but causes an 8.7fold reduction in the ability to modify amikacin
Escherichia coli
2.3.1.82
S119L
AAC(6')-IIa S119L. The AAC(6')-IIa protein modifies gentamicin C1 at 10.1% relative to sisomicin, however it shows low activity towards amikacin, 4.1% relative to sisomycin. The mutation results in a 4.8fold reduction in the acetylation of gentamicin C1, but causes an 2fold increase in the ability to modify amikacin
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2.3.1.82
additional information
Escherichia coli
involved in resistance to antibiotics
?
-
-
?
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2.3.1.82
acetyl-CoA + 2'-N-ethylnetilmicin
-
636903
Escherichia coli
CoA + N6'-acetyl-2'-N-ethylnetilmicin
-
-
-
?
2.3.1.82
acetyl-CoA + amikacin
the AAC(6')-Ib protein is unable to efficiently modify gentamicin C1, 1.7% relative activity to sisomicin, however it is capable of modifying amikacin, 65.5% relative activity to sisomycin. The mutant enzyme AAC(6')-Ib L119S shows a 2.8fold increase in acetylation of gentamicin C1, but a 8.7fold reduction in the ability to modify amikacin. The AAC(6')-IIa protein modifies gentamicin C1 at 10.1% relative activity to sisomicin, however it shows low activity towards amikacin, 4.1% activity relative to sisomycin. The mutation AAC(6')-IIa S119L results in a 4.8fold reduction in the acetylation of gentamicin C1, but causes an 2fold increase in the ability to modify amikacin
636903
Escherichia coli
CoA + N6'-acetylamikacin
-
-
-
?
2.3.1.82
acetyl-CoA + gentamicin C1
the AAC(6')-Ib protein is unable to efficiently modify gentamicin C1, 1.7% relative activity to sisomicin, however it is capable of modifying amikacin, 65.5% relative activity to sisomycin. The mutant enzyme AAC(6')-Ib L119S shows a 2.8fold increase in acetylation of gentamicin C1, but a 8.7fold reduction in the ability to modify amikacin. The AAC(6')-IIa protein modifies gentamicin C1 at 10.1% relative activity to sisomicin, however it shows low activity towards amikacin, 4.1% activity relative to sisomycin. The mutation AAC(6')-IIa S119L results in a 4.8fold reduction in the acetylation of gentamicin C1, but causes an 2fold increase in the ability to modify amikacin
636903
Escherichia coli
CoA + N6'-acetylgentamicin C1
-
-
-
?
2.3.1.82
acetyl-CoA + gentamicin C1a
poor substrate, AAC-(6')-II type
636903
Escherichia coli
CoA + N6'-acetylgentamicin C1a
-
-
-
?
2.3.1.82
acetyl-CoA + sisomicin
-
636903
Escherichia coli
CoA + N6'-acetylsisomicin
-
-
-
?
2.3.1.82
acetyl-CoA + tobramycin
-
636903
Escherichia coli
CoA + N6'-acetyltobramycin
-
-
-
?
2.3.1.82
additional information
a single amino acid, Leu119 in AAC(6')-Ib and S119 in AAC(6')-IIa, is largely responsible for determining the specificity of the AAC(6')-Ib and AAC(6')-IIa proteins. Changing this amino acid in either the AAC(6')-Ib or the AAC(6')-IIa protein results in a dramatic change in substrate specificity
636903
Escherichia coli
?
-
-
-
?
2.3.1.82
additional information
involved in resistance to antibiotics
636903
Escherichia coli
?
-
-
-
?