Structure and Function of APH(4)-Ia, a Hygromycin B Resistance Enzyme

Stogios, P.J.; Shakya, Tushar; Evdokimova, E.; Savchenko, Alexei; Wright, Gerard D.

doi:10.1074/jbc.m110.194266

Cited by 32 publications

(23 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Apart from the members of the APH(2ЈЈ) family, there is only one other APH enzyme that has been shown to unequivocally use GTP. APH(4)-Ia is able to use both GTP and ATP in the phosphorylation of hygromycin, albeit with a 5-fold preference for the latter (38). Although the structure of APH(4)-Ia has no nucleotide bound, it does have a water molecule bound in the L3/L5 pocket, and modeling of a GTP into the binding site (data not shown) suggests that a guanine nucleotide could readily be bound in a productive manner.…”

Section: Resultsmentioning

confidence: 99%

Aminoglycoside 2′′-Phosphotransferase IIIa (APH(2′′)-IIIa) Prefers GTP over ATP

Smith

Tóth

Frase

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Section: Resultsmentioning

confidence: 99%

Aminoglycoside 2′′-Phosphotransferase IIIa (APH(2′′)-IIIa) Prefers GTP over ATP

Smith

Tóth

Frase

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

“…The ribosome-binding ability of HgB is completely lost because of enzymatic phosphorylation at the 4-OH of its hyosamine moiety [30,31]. Furthermore, a synergistic relationship was observed not only between PMB and HgB but also between PMB and geneticin, which can similarly inhibit the protein synthetic reaction in eukaryotic organisms.…”

Section: Discussionmentioning

confidence: 82%

Generation of Novel Fungicidal Activity by the Combined Actions of Hygromycin B and Polymyxin B

Yutani¹,

Ogita²,

Fujita³

et al. 2013

LSMR

View full text Add to dashboard Cite

Abstract-The aminoglycoside antibiotic hygromycin B (HgB) has been widely used in veterinary medicine and in cell culture selection. HgB kills bacteria, fungi, and higher eukaryotic cells by inhibiting protein synthesis. A marked synergistic relationship was observed between HgB and the bactericidal antibiotic polymyxin B (PMB) in their ability to cause cell death of Saccharomyces cerevisiae, but this was not observed when S. cerevisiae cells were treated with PMB and a different aminoglycoside selective for prokaryotic organisms. However, the combined lethal actions of HgB and PMB did not depend on the inhibition of 80S ribosomal protein synthesis, even if the ribosome-binding OH of HgB is similarly involved in its PMB-dependent fungicidal activity. Our findings could have implications for improving the use of HgB in veterinary medicine.

show abstract

“…The sequence for residues 7 to 219 of VatA was subcloned in the Gateway-compliant ligation-independent cloning vector p15Tv-LIC (13) leading to an N-terminal His 6 -tagged protein with a tobacco etch virus (TEV) protease cleavage site between the tag and residue 7 of VatA. The protein was expressed and purified as described for APH(4)-Ia (14). The molecular weights in solution were verified by size exclusion chromatography by loading 10 to 30 l of 60 and 30 mg/ml VatA and VatD protein, respectively, onto a Superdex 200 10/ 300 GL size exclusion column (GE Healthcare) using the running buffer (0.3 M NaCl, 50 mM HEPES [pH 7.5], 0.5 mM tris [2-carboxyethyl]phosphine).…”

Section: Methodsmentioning

confidence: 99%