Hydrolysis of ATP by aminoglycoside 3'-phosphotransferases: an unexpected cost to bacteria for harboring an antibiotic resistance enzyme.
Aminoglycoside 3'-phosphotransferases (APH(3')s) are common bacterial resistance enzymes to aminoglycoside antibiotics. These enzymes transfer the gamma-phosphoryl group of ATP to the 3'-hydroxyl of the antibiotics, whereby the biological activity of the drugs is lost. Pre-steady-state and steady-state kinetics with two of these enzymes from Gram-negative bacteria, APH(3')-Ia and APH(3')-IIa, were performed. It is demonstrated that these enzymes in both ternary and binary complexes facilitate an ATP hydrolase activity (ATPase), which is competitive with the transfer of phosphate to the antibiotics. Because these enzymes are expressed constitutively in resistant bacteria, the turnover of ATP is continuous during the lifetime of the organism both in the absence and the presence of aminoglycosides. Concentrations of the enzyme in vivo were determined, and it was estimated that in a single generation of bacterial growth there exists the potential that this activity would consume as much as severalfold of the total existing ATP. Studies with bacteria harboring the aph(3')-Ia gene revealed that bacteria are able to absorb the cost of this ATP turnover, as ATP is recycled. However, the cost burden of this adventitious activity manifests a selection pressure against maintenance of the plasmids that harbor the aph(3')-Ia gene, such that approximately 50% of the plasmid is lost in 1500 bacterial generations in the absence of antibiotics. The implication is that, in the absence of selection, bacteria harboring an enzyme that catalyzes the consumption of key metabolites could experience the loss of the plasmid that encodes for the given enzyme.
Kim
Choonkeun
C
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
Cha
Joo Young
JY
Yan
Honggao
H
Vakulenko
Sergei B
SB
Mobashery
Shahriar
S
D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000617:Aminoglycosides / Q000737:chemistry; D001667:Binding, Competitive; D002384:Catalysis; D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004351:Drug Resistance; D004926:Escherichia coli / Q000378:metabolism; D006867:Hydrolases / Q000737:chemistry; D006868:Hydrolysis; D019868:Kanamycin Kinase / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D013997:Time Factors
D000617:Aminoglycosides; D017931:DNA Primers; D000255:Adenosine Triphosphate; D004247:DNA; D019868:Kanamycin Kinase; D006867:Hydrolases; D000251:Adenosine Triphosphatases