Literature summary for 2.7.1.190 extracted from

Caldwell, S.J.; Berghuis, A.M.
Plasticity of Aminoglycoside Binding to Antibiotic Kinase APH(2'')-Ia (2018), Antimicrob. Agents Chemother., 62, e00202-18 .

Search for more literature for 2.7.1.190

Crystallization (Commentary)

Crystallization (Comment)	Organism
enzyme APH(2'')-Ia crystals, grown with guanosine-beta,gamma-imidotriphosphate (GMPPNP) and a saturating concentration of magnesium, are soaked with ribostamycin, hanging drop vapour diffusion method, mixing of 0.001 ml ofp protein solution with 0.001 ml of reservoir solution containing 100 mM HEPES, pH 7.5, 120 mM MgCl2, 10% PEG 3350, and 10% glycerol, preincubation of the enzyme with 3 mM GMPPNP and 6 mM MgCl2, X-ray diffraction structure determination and analysis at 2.20-2.60 A resolution, model building	Enterococcus casseliflavus

Crystallization (Comment)

Organism

enzyme APH(2'')-Ia crystals, grown with guanosine-beta,gamma-imidotriphosphate (GMPPNP) and a saturating concentration of magnesium, are soaked with ribostamycin, hanging drop vapour diffusion method, mixing of 0.001 ml ofp protein solution with 0.001 ml of reservoir solution containing 100 mM HEPES, pH 7.5, 120 mM MgCl2, 10% PEG 3350, and 10% glycerol, preincubation of the enzyme with 3 mM GMPPNP and 6 mM MgCl2, X-ray diffraction structure determination and analysis at 2.20-2.60 A resolution, model building

Enterococcus casseliflavus

Protein Variants

Protein Variants	Comment	Organism
S376N	a clinically identified naturally occuring S376N mutation of APH(2'')-Ia elevates resistance to N1-substituted aminoglycosides but eliminates modification of nonsubstituted compounds. Mutation of serine 376 to asparagine does not lead to substantial rearrangements in the aminoglycoside binding site. In fact, the addition of the larger asparagine residue in place of serine 376 creates an obstruction that prevents binding in the neamine binding pocket. As a result, any compounds that bind using the neamine rings are blocked from the antibiotic binding site of APH(2x02)-Ia. But the mutation remains compatible with one of the alternate binding modes of amikacin, which does not use this site to interact with the enzyme	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.
GTP + dibekacin	Enterococcus casseliflavus	-	GDP + dibekacin 2''-phosphate	-	?
GTP + gentamicin	Enterococcus casseliflavus	-	GDP + gentamicin 2''-phosphate	-	?
GTP + kanamycin	Enterococcus casseliflavus	-	GDP + kanamycin 2''-phosphate	-	?
GTP + lividomycin	Enterococcus casseliflavus	-	GDP + lividomycin 2''-phosphate	-	?
GTP + neomycin	Enterococcus casseliflavus	-	GDP + neomycin 2''-phosphate	-	?
GTP + ribostamycin	Enterococcus casseliflavus	-	GDP + ribostamycin 2''-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Enterococcus casseliflavus	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
GTP + dibekacin	-	Enterococcus casseliflavus	GDP + dibekacin 2''-phosphate	-	?
GTP + gentamicin	-	Enterococcus casseliflavus	GDP + gentamicin 2''-phosphate	-	?
GTP + kanamycin	-	Enterococcus casseliflavus	GDP + kanamycin 2''-phosphate	-	?
GTP + lividomycin	-	Enterococcus casseliflavus	GDP + lividomycin 2''-phosphate	-	?
GTP + lividomycin	enzyme binding structure analysis, overview	Enterococcus casseliflavus	GDP + lividomycin 2''-phosphate	-	?
GTP + neomycin	-	Enterococcus casseliflavus	GDP + neomycin 2''-phosphate	-	?
GTP + ribostamycin	-	Enterococcus casseliflavus	GDP + ribostamycin 2''-phosphate	-	?
additional information	most aminoglycosides are based upon a neamine core, i.e. a central 2-deoxystreptamine (2-DOS) ring with an aminohexose sugar linked to 2-DOS at the 4-position. These rings form the minimal active element of an aminoglycoside antibiotic, and two subclasses of aminoglycosides are formed from elaboration of this scaffold. Addition of rings to the 5- and 6-positions of 2-DOS differentiate aminoglycosides into the 4,5-disubstituted (ribostamycin, neomycin, and lividomycin) and 4,6-disusbstituted (kanamycin, gentamicin, and dibekacin) groups	Enterococcus casseliflavus	?	-	-

Synonyms

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

Cofactor

Cofactor	Comment	Organism	Structure
GTP	-	Enterococcus casseliflavus

General Information

General Information	Comment	Organism
malfunction	if the enzyme's binding mode is made impossible because of additional substitutions to the standard 4,5- or 4,6-disubstituted aminoglycoside architecture, as in lividomycin A or the N1-substituted aminoglycosides, it is still possible for these aminoglycosides to bind to the antibiotic binding site by using alternate binding modes, which explains the low rates of noncanonical phosphorylation activities seen in enzyme assays. A clinically observed arbekacin-resistant mutant of APH(2'')-Ia reveals an altered aminoglycoside binding site that can stabilize an alternative binding mode for N1-substituted aminoglycosides. This mutation may alter and expand the aminoglycoside resistance spectrum of the wild-type enzyme in response to developed aminoglycosides	Enterococcus casseliflavus
additional information	APH(2'')-Ia maintains a preferred mode of binding aminoglycosides by using the conserved neamine rings when possible, with flexibility that allows it to accommodate additional rings	Enterococcus casseliflavus
physiological function	the APH(2'')-Ia aminoglycoside resistance enzyme forms the C-terminal domain of the bifunctional AAC(6')-Ie/APH(2'')-Ia enzyme and confers high-level resistance to natural 4,6-disubstituted aminoglycosides. The enzyme can phosphorylate 4,5-disubstituted compounds and aminoglycosides with substitutions at the N1 position	Enterococcus casseliflavus