Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1984 Jun;25(6):754–759. doi: 10.1128/aac.25.6.754

Purification and characterization of aminoglycoside-modifying enzymes from Staphylococcus aureus and Staphylococcus epidermidis.

K Ubukata, N Yamashita, A Gotoh, M Konno
PMCID: PMC185635  PMID: 6331299

Abstract

Several strains of Staphylococcus aureus and Staphylococcus epidermidis, exhibiting characteristic resistance patterns to aminoglycoside antibiotics, were examined. The aminoglycoside-modifying enzymes from these strains were purified by DEAE-Sephadex A-50 chromatography, affinity chromatography, and Sephadex G-100 gel filtration. Three enzymes, a 3'-phosphotransferase III (molecular weight, 31,000; pI 4.1), a bifunctional enzyme having 6'-acetyltransferase and 2"-phosphotransferase (molecular weight, 56,000; pI 4.1) activity, and a 4'4"-adenylytransferase (molecular weight, 34,000; pI 4.7), were isolated from crude extracts of the resistant strains. Aminoglycoside-modifying enzymes with identical enzymatic properties derived from S. aureus and S. epidermidis were also immunologically identical.

Full text

PDF
755

Images in this article

757Image
on p.757
757Image
on p.757

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Cohen M. L., Wong E. S., Falkow S. Common R-plasmids in Staphylococcus aureus and Staphylococcus epidermidis during a nosocomial Staphylococcus aureus outbreak. Antimicrob Agents Chemother. 1982 Feb;21(2):210–215. doi: 10.1128/aac.21.2.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Courvalin P., Davies J. Plasmid-medicated aminoglycoside phosphotransferase of broad substrate range that phosphorylates amikacin. Antimicrob Agents Chemother. 1977 Apr;11(4):619–624. doi: 10.1128/aac.11.4.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dowding J. E. Mechanisms of gentamicin resistance in Staphylococcus aureus. Antimicrob Agents Chemother. 1977 Jan;11(1):47–50. doi: 10.1128/aac.11.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Forbes B. A., Schaberg D. R. Transfer of resistance plasmids from Staphylococcus epidermidis to Staphylococcus aureus: evidence for conjugative exchange of resistance. J Bacteriol. 1983 Feb;153(2):627–634. doi: 10.1128/jb.153.2.627-634.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Haas M. J., Dowding J. E. Aminoglycoside-modifying enzymes. Methods Enzymol. 1975;43:611–628. doi: 10.1016/0076-6879(75)43124-x. [DOI] [PubMed] [Google Scholar]
  7. Jaffe H. W., Sweeney H. M., Nathan C., Weinstein R. A., Kabins S. A., Cohen S. Identity and interspecific transfer of gentamicin-resistance plasmids in Staphylococcus aureus and Staphylococcus epidermidis. J Infect Dis. 1980 Jun;141(6):738–747. doi: 10.1093/infdis/141.6.738. [DOI] [PubMed] [Google Scholar]
  8. Jaffe H. W., Sweeney H. M., Weinstein R. A., Kabins S. A., Nathan C., Cohen S. Structural and phenotypic varieties of gentamicin resistance plasmids in hospital strains of Staphylococcus aureus and coagulase-negative staphylococci. Antimicrob Agents Chemother. 1982 May;21(5):773–779. doi: 10.1128/aac.21.5.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kono M., Sasatsu M., Ubukata K., Konno M., Fujii R. New plasmid (pTU512), mediating resistance to penicillin, erythromycin, and kanamycin, from clinical isolates of Staphylococcus aureus. Antimicrob Agents Chemother. 1978 Apr;13(4):691–694. doi: 10.1128/aac.13.4.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  11. Le Goffic F., Martel A., Capmau M. L., Baca B., Goebel P., Chardon H., Soussy C. J., Duval J., Bouanchaud D. H. New plasmid-mediated nucleotidylation of aminoglycoside antibiotics in Staphlococcus aureus. Antimicrob Agents Chemother. 1976 Aug;10(2):258–264. doi: 10.1128/aac.10.2.258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Le Goffic F. The resistance of S aureus to aminoglycoside antibiotics and pristinamycins in France in 1976-1977. Jpn J Antibiot. 1977 Dec;30 (Suppl):286–291. [PubMed] [Google Scholar]
  13. Martel A., Moreau N., Capmau M. L., Soussy C. J., Duval J. 2"-O-phosphorylation of gentamicin components by a Staphylococcus aureus strain carrying a plasmid. Antimicrob Agents Chemother. 1977 Jul;12(1):26–30. doi: 10.1128/aac.12.1.26. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matsuhashi Y., Sawa T., Takeuchi T., Umezawa H. Immunological studies of aminoglycoside 3'-phosphotransferases. J Antibiot (Tokyo) 1976 Oct;29(10):1127–1128. doi: 10.7164/antibiotics.29.1127. [DOI] [PubMed] [Google Scholar]
  15. McDonnell R. W., Sweeney H. M., Cohen S. Conjugational transfer of gentamicin resistance plasmids intra- and interspecifically in Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother. 1983 Jan;23(1):151–160. doi: 10.1128/aac.23.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McGowan J. E., Jr, Terry P. M., Huang T. S., Houk C. L., Davies J. Nosocomial infections with gentamicin-resistant Staphylococcus aureus: plamid analysis as an epidemiologic tool. J Infect Dis. 1979 Dec;140(6):864–872. doi: 10.1093/infdis/140.6.864. [DOI] [PubMed] [Google Scholar]
  17. Porthouse A., Brown D. F., Smith R. G., Rogers T. Gentamicin resistance in Staphylococcus aureus. Lancet. 1976 Jan 3;1(7949):20–21. doi: 10.1016/s0140-6736(76)92912-3. [DOI] [PubMed] [Google Scholar]
  18. Santanam P., Kayser F. H. Purification and characterization of an aminoglycoside inactivating enzyme from Staphylococcus epidermidis FK109 that nucleotidylates the 4'- and 4''-hydroxyl groups of the aminoglycoside antibiotics. J Antibiot (Tokyo) 1978 Apr;31(4):343–351. doi: 10.7164/antibiotics.31.343. [DOI] [PubMed] [Google Scholar]
  19. Santanam P., Kayser F. H. Tobramycin adenylyltransferase: a new aminoglycoside-inactivating enzyme from Staphylococcus epidermidis. J Infect Dis. 1976 Aug;134 (Suppl):S33–S39. doi: 10.1093/infdis/134.supplement_1.s33. [DOI] [PubMed] [Google Scholar]
  20. Scott D. F., Wood D. O., Brownell G. H., Carter M. J., Best G. K. Aminoglycoside modification by gentamicin-resistant isolates of Staphylococcus aureus. Antimicrob Agents Chemother. 1978 Apr;13(4):641–644. doi: 10.1128/aac.13.4.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Umezawa H., Yamamoto H., Yagisawa M., Kondo S., Takeuchi T. Letter: Kanamycin phosphotransferase. I. Mechanism of cross resistance between kanamycin and lividomycin. J Antibiot (Tokyo) 1973 Jul;26(7):407–411. doi: 10.7164/antibiotics.26.407. [DOI] [PubMed] [Google Scholar]
  22. Vogel L., Nathan C., Sweeney H. M., Kabins S. A., Cohen S. Infections due to gentamicin-resistant Staphylococcus aureus strain in a nursery for neonatal infants. Antimicrob Agents Chemother. 1978 Mar;13(3):466–472. doi: 10.1128/aac.13.3.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weinstein R. A., Kabins S. A., Nathan C., Sweeney H. M., Jaffe H. W., Cohen S. Gentamicin-resistant staphylococci as hospital flora: epidemiology and resistance plasmids. J Infect Dis. 1982 Mar;145(3):374–382. doi: 10.1093/infdis/145.3.374. [DOI] [PubMed] [Google Scholar]
  24. Wood D. O., Carter M. J., Best G. K. Plasmid-mediated resistance to gentamicin in Staphylococcus aureus. Antimicrob Agents Chemother. 1977 Oct;12(4):513–517. doi: 10.1128/aac.12.4.513. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES