Literature summary for 2.7.1.190 extracted from

Kaplan, E.; Guichou, J.F.; Chaloin, L.; Kunzelmann, S.; Leban, N.; Serpersu, E.H.; Lionne, C.
Aminoglycoside binding and catalysis specificity of aminoglycoside 2''-phosphotransferase IVa A thermodynamic, structural and kinetic study (2016), Biochim. Biophys. Acta, 1860, 802-813 .

Search for more literature for 2.7.1.190

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3). The catalytic activity of APH(2'')-IVa is independent of the presence of the 6His-tag	Enterococcus casseliflavus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme in complex with substrate G418 and sisomicin, mixing 0.001 ml of 6 mg/ml protein and 2.5 mM aminoglycoside in 50 mM HEPES, pH 7.5, and 10 mM MgCl2 with 0.001 ml of reservoir solution containing 12% PEG 3350 w/v, and 50-75 mM ammonium citrate, pH 7.4-7.9, 18°C, X-ray diffraction structure determination and analysis at 3.05 A and 2.35 A resolution, respectively, molecular replacement using chain A of the homologous APH(2'')-IVa-ADP complex (PDB ID 4N57) after removal of the ligand as search model, modeling	Enterococcus casseliflavus

Crystallization (Comment)

Organism

purified enzyme in complex with substrate G418 and sisomicin, mixing 0.001 ml of 6 mg/ml protein and 2.5 mM aminoglycoside in 50 mM HEPES, pH 7.5, and 10 mM MgCl2 with 0.001 ml of reservoir solution containing 12% PEG 3350 w/v, and 50-75 mM ammonium citrate, pH 7.4-7.9, 18°C, X-ray diffraction structure determination and analysis at 3.05 A and 2.35 A resolution, respectively, molecular replacement using chain A of the homologous APH(2'')-IVa-ADP complex (PDB ID 4N57) after removal of the ligand as search model, modeling

Enterococcus casseliflavus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state and transient kinetics	Enterococcus casseliflavus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Enterococcus casseliflavus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
GTP + gentamicin	Enterococcus casseliflavus	-	GDP + gentamicin 2''-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Enterococcus casseliflavus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) bynickel affinity chromatography, ultrafiltration, and gel filtration	Enterococcus casseliflavus

Reaction

Reaction	Comment	Organism	Reaction ID
GTP + gentamicin = GDP + gentamicin 2''-phosphate	the reaction of APH(2'')-IVa proceeds by a Bi-Bi mechanism in which aminoglycoside and ATP or GTP bind randomly	Enterococcus casseliflavus	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
GTP + G418	-	Enterococcus casseliflavus	GDP + G418 2''-phosphate	-	?
GTP + gentamicin	-	Enterococcus casseliflavus	GDP + gentamicin 2''-phosphate	-	?
GTP + sisomicin	-	Enterococcus casseliflavus	GDP + sisomicin 2''-phosphate	-	?

Synonyms

Synonyms	Comment	Organism
aminoglycoside 2''-phosphotransferase IVa	-	Enterococcus casseliflavus
APH(2'')-IVa	-	Enterococcus casseliflavus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Enterococcus casseliflavus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Enterococcus casseliflavus

Cofactor

Cofactor	Comment	Organism	Structure
GTP	-	Enterococcus casseliflavus