Backbone resonance assignments of a promiscuous aminoglycoside antibiotic resistance enzyme; the aminoglycoside phosphotransferase(3')-IIIa

Serpersu E. H., ÖZEN C., Norris A. L., Steren C., Whittemore N.

BIOMOLECULAR NMR ASSIGNMENTS, vol.4, no.1, pp.9-12, 2010 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 4 Issue: 1
  • Publication Date: 2010
  • Doi Number: 10.1007/s12104-009-9195-z
  • Journal Name: BIOMOLECULAR NMR ASSIGNMENTS
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.9-12
  • Middle East Technical University Affiliated: Yes

Abstract

The aminoglycoside phosphotransferase(3')-IIIa (APH) is a promiscuous enzyme and renders a large number of structurally diverse aminoglycoside antibiotics useless against infectious bacteria. A remarkable property of this similar to 31 kDa enzyme is in its unusual dynamic behavior in solution; the apo-form of the enzyme exchanges all of its backbone amide protons within 15 h of exposure to D (2) O while aminoglycoside-bound forms retain similar to 40% of the amide protons even after > 90 h of exposure. Moreover, the number of observable peaks and their dispersion in HSQC spectra varies with each aminoglycoside, rendering the resonance assignments very challenging. Therefore, the binary APH-tobramycin complex, which shows the largest number of well-resolved peaks, was used for the backbone resonance assignments (C alpha, C, N, H, and some C beta) of this protein (BMRB-16337).