Kinetic studies of aminoglycoside acetyltransferase and phosphotransferase from Staphylococcus aureus RPAL. Relationship between the two activities

Eur J Biochem. 1983 Jul 1;133(3):515-21. doi: 10.1111/j.1432-1033.1983.tb07494.x.


In the Staphylococcus aureus strain harbouring the plasmid RPAL, the resistance to aminoglycoside antibiotics results from two inactivating reactions catalyzed by a 6'-N-aminoglycoside acetyltransferase and a 2"-O-amino-glycoside phosphotransferase. These enzymes are copurified with a constant ratio between the two activities, the purification process consisting in affinity chromatography, native electrophoresis and gel exclusion chromatography. The kinetic mechanisms of each activity have been determined from studies of initial velocities, as well as product and dead-end inhibitions. Both activities follow a random rapid equilibrium mechanism. The substrates and cofactors of one reaction have been tested as effectors of the other reaction. No interaction between the two activities has been observed. However, the GTP cofactor of phosphotransferase protects, at weak concentrations, the acetyltransferase against thermal inactivation, which suggests that the two activities may be associated.

Publication types

  • Comparative Study

MeSH terms

  • Acetyltransferases / antagonists & inhibitors
  • Acetyltransferases / isolation & purification*
  • Bacterial Proteins / isolation & purification
  • Chemical Phenomena
  • Chemistry
  • Kanamycin Kinase
  • Kinetics
  • Phosphotransferases / antagonists & inhibitors*
  • Staphylococcus aureus / enzymology*
  • Substrate Specificity
  • Temperature


  • Bacterial Proteins
  • Acetyltransferases
  • aminoglycoside acetyltransferase
  • Phosphotransferases
  • Kanamycin Kinase